THREE
The consulting room of Dr. Colenso Ridgeon, KBE, was, in the spring of 1903, located on the second floor of an unremarkable building in London’s Marylebone neighborhood, indistinguishable from a hundred other Victorian apartments. In addition to an overstuffed couch and chairs, the green-wallpapered room featured a marble-topped console table with gilt legs ending in the claws of a sphinx; a mirror so covered with paintings of palms, lilies, and other plants that it was no longer useful as a reflecting surface; and a writing table, on which a microscope, test tubes, and a small alcohol stove fought for space with piles of papers and journals. Like the doctor himself, the room made some concession to modernity—it had replaced gas lighting with electricity—but otherwise looked, in the first years of the twentieth century, almost exactly as it had in the middle of the nineteenth: a very model of bourgeois solidity.
If Dr. Ridgeon’s place of work was conservative, the work he did there was positively revolutionary. So revolutionary, in fact, that in June 1903 he received a knighthood for it. After thirty years as a practicing physician, Dr. Ridgeon had discovered a cure for tuberculosis, an historic achievement if anything was.
You will, however, search in vain through any history of medicine looking for him, or his discovery. Colenso Ridgeon was and remains a creature of the imagination whose genius (and taste in furniture) was described, and his knighthood awarded, in the first act of a 1906 play entitled The Doctor’s Dilemma, written by the Irish iconoclast George Bernard Shaw.
Shaw was a Fabian Socialist, a sometime atheist, an antivivisectionist, and an Irish patriot, but his real passion was satire, and he had written the play to skewer the medical establishment of his day. Debating the “dilemma”—whether Ridgeon should use his tuberculosis cure to treat an honorable but inconsequential friend, or a deeply immoral but brilliant artist, a choice complicated by the doctor’s lust for the painter’s wife—are a group of physicians with what might charitably be called blinkered approaches to the healing profession. A onetime schoolmate of Dr. Ridgeon, Dr. Leo Schutzmacher, offers the two-word secret to his successful practice—“Cure Guaranteed”—while confiding, “You see, most people get well all right if they are careful and you give them a little sensible advice.” Cutler Walpole is a frighteningly eager surgeon who believes all disease is a version of blood poisoning, which is invariably cured by the removal of an entirely invented organ he calls the “nuciform sac.” Sir Ralph Bloomfield Bonington, having read a page or two from Koch and Pasteur, treats each of his patients with the most dangerous microbes he can find, expecting thereby to promote a natural cure. “Drugs are a delusion. Find the germ of the disease; prepare from it a suitable anti-toxin; inject it three times a day quarter of an hour before meals; and what is the result? The phagocytes* are stimulated; they devour the disease; and the patient recovers.” Of them all, only the now-retired Sir Patrick Cullen escapes Shaw’s barbs, pointing out to Ridgeon that, despite his newfound cure, “I’ve known over thirty men that found out how to cure consumption. Why do people go on dying of it?”
For a satire to work at all, its targets must be familiar to its audience, and so it was with The Doctor’s Dilemma. Every one of the London playgoers who attended its premiere would have known physicians who reflexively removed each of their patient’s tonsils or appendix irrespective of their presenting symptoms; or those who monomaniacally prescribed antitoxins or antiserums for everything from skin lesions to cancer. And they would have recognized the model that Shaw used for Colenso Ridgeon: the nation’s most famous physician, Dr. Almroth Edward Wright.
In 1906, Wright, a friend and occasional debating opponent of Shaw, was forty-five years old, and already frequently referred to in the press as “Britain’s Pasteur.” He had come to medicine by a somewhat circuitous route; before graduating as a physician from Trinity College, Dublin, in 1883—Wright was partly Irish by descent—he had already taken First-Class Honours in modern literature, and would later read “Jurisprudence and International Law with a View to the Bar.” From 1895 on, though, he was completely devoted to medicine, and his particular area of interest was the discovery of an immunization against typhoid fever, one of the deadliest diseases in history.
Variously blamed for the fifth century B.C.E. Plague of Athens, the destruction of the English colony of Jamestown in the seventeenth century, and the deaths of tens of thousands of American Civil War soldiers, typhoid fever is one of a dozen different diseases caused by a strain of bacteria from the genus Salmonella, specifically S. typhi.* Typhoid fever is a killer, but not a sudden one; a typical infection lasts up to a month, beginning with a characteristic low-grade fever and slowed heartbeat. As the fever rises, the heart continues to slow, frequently accompanied by delirium. The bacteria reproduce rapidly, causing the host’s abdomen to distend and severe diarrhea to follow. Internal organs like the spleen and liver enlarge. Intestines hemorrhage, and sometimes perforate, spreading infection to the internal membrane: the peritoneum. Unless the victim’s immune system is successful in fighting off these sequential attacks, death results; mortality can run as high as 25 percent in untreated typhoid fever.
The destructiveness of any infectious disease is a function of both virulence and mobility: how much damage the pathogen causes, and how easily it travels. Because S. typhi spreads by consumption of drinking water contaminated with human feces and urine, it was particularly deadly wherever large numbers of people with poor access to sanitation lived in close contact: the poorer quarters of rapidly growing nineteenth-century cities, for example. Or—even more lethally—armies in the field. During the Spanish-American War, typhoid fever killed more American soldiers than either battle wounds or even the feared viral disease known as yellow fever.
For obvious reasons, then, military doctors were particularly concerned about the disease, none more so than Wright, whose first job after becoming a physician was at the Army Medical School at Netley, near Southampton. There, in 1895, he developed the first effective vaccine against typhoid, inoculating subjects not with a weakened version of the S. typhi bacterium (this was the method tested by Robert Koch), but, far more safely and effectively, a dead one. He tested it first on himself, then on fifteen volunteers, and finally on a regiment’s worth of British soldiers headed to India. It was Wright’s first, and greatest, triumph: Of nearly three thousand subjects, only ten contracted the disease.
Despite this success, he was unable to persuade the conservative policymakers in Britain’s War Department to inoculate troops being sent to South Africa in 1899 to fight in the Second Boer War. As a result of their reactionary hostility to even modern medicine, even in the face of what seems its inarguable success in India, over the next three years typhoid proved more deadly to the British army than the combined efforts of the Afrikaner Transvaal Republic and the Orange Free State. At least twenty thousand British soldiers contracted the disease, and more than nine thousand of them died. Disgusted, Wright left the War Department in 1902, and moved to St. Mary’s Hospital. On Praed Street in London’s Paddington neighborhood, St. Mary’s was one of the last of London’s so-called “voluntary” hospitals, set up for the care of the working poor, and one of the first conceived of as a teaching hospital with an attached medical school. There, following the model of Pasteur and Koch, he opened a laboratory—the “Inoculation Department”—that he would direct for the next forty-five years.
In retrospect, Wright’s accomplishments at St. Mary’s never soared as high as his reputation, and his place in history has suffered in consequence. During his lifetime he was hugely famous and influential, eccentric and intimidating. In legend, at least, his memory was so remarkable that he had committed a quarter of a million lines of poetry to it. Wright was tall and striking, careless of his dress but wickedly entertaining in his speech, a brilliant raconteur and lecturer, and a public figure whose opinion was sought on issues scientific, social, and political until his death in 1947. He was also a fine and innovative experimentalist, and a master of laboratory technique. With little more than a microscope, a Bunsen burner, and a supply of rubber nipples and glass tubing, he was able to perform extraordinarily sophisticated research* in ways that remind historians of science just how much the lab once depended on the steady hand of a craftsman. The vials that Wright used to collect blood, his so-called blood capsules, were custom-made bits of glass pipette that he melted and drew in the flame of a Bunsen burner into narrow tubes that he then bent at an angle. Snipping the glass at one end provided a needle, and the curve allowed the glass straw to draw the blood by capillary action.
Credit: Wellcome Library, London
Almroth Wright, 1861–1947
Wright’s lab skills were, however, something of a two-edged sword, since they made him prone to accept his acute clinical observations as irrefutable proof. Yet he was hopeless with numbers; the original dead-cell typhoid inoculation he performed on 2,835 India-bound soldiers was almost certainly successful, but you couldn’t prove it by Wright’s statistics. According to Britain’s leading mathematical biologist, Karl Pearson, the data Wright collected were useless for concluding anything: no control groups, no attempts to show what statisticians call the “null hypothesis”—the assumption that there is no relationship between two phenomena, such as “being inoculated” and “getting typhoid.” Wright’s statistical illiteracy was likely a consequence less of his temperament than of his eccentric education, which was almost willfully deficient in practical mathematics, even by the standards of nineteenth-century Great Britain. Wright had been home tutored, and spent far more time on Latin declensions and the history of the common law than on regression analysis.* It is almost certainly that blind spot that explains his devotion to one of the great dead ends in medical history: vaccine therapy, the use of substances that activate the adaptive immune system to fight a specific disease as a therapy, rather than a preventative.
Wright was a vaccine absolutist, famously observing, “The physician of the future will be an immunizator.” The key, to Wright, was the particular character of an individual patient’s immune system, not an attack on pathogens using chemicals such as Ehrlich’s Salvarsan or, later, Domagk’s sulfanilamide. This debate—whether disease was best understood as what occurs when a healthy host encounters a pathogen, or as the consequence of what happens when an unhealthy host does so, with the latter providing evidence of a deficiency in the host’s internal environment—dates back to Pasteur, and, in some senses, remains alive today.* Convinced by the promising results of serum therapy to treat illnesses such as rabies, Wright predicted that similar techniques could be used “to exploit the uninfected tissues in favor of the infected.” Wright named this phenomenon the “opsonic mechanism.”
In explaining the wholly fictional tuberculosis cure at the heart of The Doctor’s Dilemma, Colenso Ridgeon says to Sir Patrick Cullen that “opsonin is what you butter the disease germs with to make your white blood corpuscles eat them.” Shaw was prescient. A story in the New York Times from March 31, 1907—a year after the premiere of the play—is headlined: “THE NEW HOPE FOR TUBERCULOSIS: DISCOVERY OF ‘OPSONINS’ PROMISES TO REVOLUTIONIZE MEDICINE.” The newspaper goes on to quote Wright on his discovery that opsonins do their work “by uniting with the micro-organisms, the invading germs, and rendering them more palatable, so to speak, to the white corpuscles.”*
Opsonins are real. Any molecule that enhances the way white blood cells ingest and kill invading pathogens is, technically, an opsonin, as are those that activate the complement that is part of the innate immune system. Opsonic therapy, however, never lived up to its initial promise; nor did Almroth Wright. “Britain’s Pasteur” almost certainly saved hundreds of thousands of lives during the First World War; the British army that fought on the western front was given Wright’s typhoid inoculation, and only twelve hundred soldiers died of it, out of more than two million. He performed heroically during the war itself, demonstrating the limits of antiseptic pastes and liquids like Lister’s carbolic acid to treat battlefield wounds—carbolic acid didn’t just attack pathogens, but the immune system’s leukocytes as well—and the dangers of airtight bandages, which encouraged the growth of nasty, gangrene-causing bacteria like Clostridium perfringens that thrive in anaerobic environments. Nonetheless, he is now mostly remembered as Britain’s leading opponent of women’s suffrage. And, of course, as the inspiration for Colenso Ridgeon, Shaw’s dilemma-facing doctor.
This scants Wright’s real legacy: the Inoculation Department he founded and ran for decades at St. Mary’s, and that he made into an incubator for the next generation of antibacterial researchers. One of his subordinates there, who followed him to France, was Leonard Colebrook.
Another was a Scottish physician named Alexander Fleming.
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When Fleming joined Almroth Wright at the Inoculation Department in 1906, the then twenty-five-year-old physician was a promising if not yet accomplished researcher. He was also a sort of anti-Wright—where Wright was tall and physically imposing, Fleming was short and slender; Wright had a mustache that made walruses envious, Fleming was clean-shaven; and while Wright was never happier than when speaking publicly, Fleming was so self-effacing that students had to strain to hear his lectures. He had graduated with distinction both from the Royal Polytechnic Institution (now the University of Westminster) and from St. Mary’s Hospital Medical School, where he had trained as a surgeon before discovering a talent for experimental research. In 1909, Fleming designed a new test for syphilis that required less blood, and was more effective, than the eponymous diagnostic invented three years before by the German bacteriologist August Paul von Wassermann. The following year, Fleming began working with Leonard Colebrook to investigate the properties of Ehrlich’s magic bullets: Salvarsan and Neosalvarsan.
When the First World War broke out, Fleming and Colebrook accompanied Wright to France, where the core of St. Mary’s Inoculation Department joined the British military hospital at Boulogne-sur-Mer. That hospital, one of several built to accommodate the huge number of casualties from the first Battle of Ypres—the same battle in which Gerhard Domagk received his wound—would prove a remarkably productive research facility, even without considering the circumstances under which it had been established. While working at the hospital, the St. Mary’s team discovered that the standard of care for wounds—antiseptic ointments and airtight bandages—actually promoted infections rather than preventing them. As Fleming recognized, the cause was the variety of bacteria that grow even in the absence of oxygen,* particularly under the skin’s surface. And those bacteria were, literally, everywhere, even after exposure to powerful antiseptics. It took some experimental skill to demonstrate why.
As he described in a now-classic paper written for the Lancet, Fleming exposed two sets of glass tubes to a highly concentrated bacterial soup. One set was left whole, while the other was broken to create a ragged edge that would simulate a battlefield wound. After both were washed with antiseptics, the unbroken test tubes were completely disinfected, but the bacteria in the broken tube’s hidden recesses stubbornly reappeared, even after washing in carbolic acid. Fleming had demonstrated experimentally why even unbloodied uniforms from soldiers with supposedly disinfected wounds remained rife with pathogens. Dangerous ones. Fifteen percent of battlefield woundscontained staph, 30 percent tetanus, 40 percent strep . . . and 90 percent were infected with the gangrene-causing C. perfringens.
In November 1918, the First World War ended. For Fleming, now returned to St. Mary’s, the war against pathogenic bacteria was just getting started. He had acquired a more sophisticated understanding of the resourcefulness of his opponents, but was no closer to victory over them until, in 1922, he made his first improbably accidental discovery. As his laboratory assistant, V. D. Allison, later recalled, Fleming:
. . . was busy one evening cleaning up several Petri dishes which had been lying on the bench for perhaps ten days or a fortnight. As he took up one of the dishes in his hand, he looked at it for a long time, showed it to me, and said: “This is interesting.” . . . It was covered with large yellow colonies which appeared to me to be obvious contaminants. But the remarkable fact was that there was a wide area in which there were no organisms. . . . Fleming explained that this particular dish was one to which he had added a little of his own nasal mucus, when he had happened to have a cold. The mucus was in the middle of the zone containing no colony. The idea at once occurred to him that there must be something in the mucus that dissolved or killed the microbes. . . .
Fleming named the substance found in his mucus lysozyme: the first purely organic substance shown to have antibacterial properties. However, the unlikelihood of the discovery as reported seems almost too much to credit. First, Fleming later revealed that the mucus had accidentally dripped from his nose onto one of the Petri dishes. Not just dripped, but dripped onto the one Petri dish that had, somehow, picked up a bacterial contaminant from a fortuitously open window . . . and, even less probably, since most bacteria (and all important pathogens) are unaffected by lysozyme, the contaminant on the twice-lucky dish would have had to be one of the few bacteria with lysozyme sensitivity. This is the laboratory equivalent of buying winning lottery tickets twice on the same day.
However improbable its discovery, lysozyme was an interesting, but relatively inconsequential compound, one that Fleming accurately recognized as an enzyme: a large molecule that increases the speed of organic chemical reactions. Some years later, it was identified as one of the components of the body’s innate immune system, whose activity works to damage bacterial cell walls. This is a nontrivial ability that offers some protection against infection, particularly in newborn children, but isn’t much use against most pathogens. The same can’t be said of Fleming’s next encounter with good fortune, which occurred some five years later.
Credit: Wellcome Library, London
Alexander Fleming, 1881–1955
The canonical story of the discovery of penicillin is eerily similar to the one describing the chance discovery of lysozyme. As Fleming later recalled, he had sloppily left Petri dishes containing staph cultures unattended on a bench in his St. Mary’s lab when he departed for vacation in August 1928. When he returned, on September 3, he found that one of the Petri dishes had been contaminated, again via a conveniently open window, this time by a fungus. The evidence was even more startling than five years earlier: Around the fungal contamination was a ring in which all the staphylococci had disappeared. Something had killed them.
For weeks, Fleming worked to cultivate the fungus, Penicillium notatum, technically a mold (molds—in Britain, “moulds”—are fungi that take the form of tiny multicellular filaments, which gives them their characteristically fuzzy appearance; unicellular fungi are yeasts). The mold was producing some substance that was deadly to the staph bacteria, a substance that Fleming first named “penicillin” in March 1929 in an article entitled “On the Antibacterial Action of Cultures of a Penicillium, with Special Reference to Their Use in the Isolation of B. Influenzae.”
So: another open window. Another accidental contamination. Another brilliant discovery illustrating the power of serendipity.
Or, perhaps not. For decades, historians and scientists have puzzled over the inconsistencies in Fleming’s account. For one thing, the famous window in Fleming’s lab (today preserved as a museum) appears to have rarely, if ever, been opened in order to avoid exactly what Fleming later claimed: the accidental introduction of a contaminant. For another, the timing seems as fuzzy as the fungus itself: The original account has the Petri dishes left alone for more than five weeks, but in 1944 Fleming himself said that the effect was observed after only two. Though Fleming’s chronology puts the discovery of the world-historic Petri dish on September 3, the first day in which he noted it in his lab notebooks is October 30.
Most tellingly of all: If the Penicillium mold had appeared after the staph colonies were well established, they would have killed it long before it could produce penicillin in the first place. Fleming could not, of course, have known this, but the way penicillin kills or degrades staphylococci is by disrupting the mechanism the bacterium uses to build new walls during the process of cell division—which means that it works only when the bacteria are dividing.*Unless the mold is present before the staph, no ring of bacterial death.
This doesn’t mean that Fleming was fraudulent, or even forgetful, either in his account of the discovery of lysozyme or of penicillin. The most appealing explanation for the discrepancies between either account and, well, logic, is something else: playfulness.
Fleming grew up in a family that was notoriously fond of cards, table tennis, and quizzes. As an adult, he was an avid player of croquet and snooker, and a skilled rifle shot (he was originally recruited to St. Mary’s as a shooter for the hospital’s famously competitive sports teams). As a golfer, he far preferred what might be called creative versions of the game, up to and including using his putter as a pool cue. He painted, too; not well, but certainly inventively: When he was a young researcher at St. Mary’s, Fleming regularly created images—Madonna and Child; the logo of St. Mary’s; the Union Jack—on Petri dishes, using agar as the canvas and microorganisms that turned different colors as they grew as pigments. (It’s worth noting that this kind of “painting” demanded an almost terrifying level of both bacteriological knowledge—which ones turn red, which green, and when—and hand-eye coordination.) One of his biographers observed: “Fleming’s natural level was indeed play. . . . ‘I play with microbes’—his often repeated description of his work—was literally true. Most of his research was a game to him and indeed most of his enjoyment came from games of all kinds.”
But while Fleming loved to play with microbes, he was almost completely at sea when it came to human interaction: a terrible conversationalist and a worse lecturer. He was painfully shy, and had no interest in discussing either his methods or results, which is why it’s at least plausible that Fleming’s penchant for games, combined with his natural reticence, persuaded him to cast the lysozyme discovery as if it had been the equivalent of drawing a winning hand at bridge.
Understanding why he would trot out such a similar story for the far more important discovery of penicillin requires some additional context. The first, and most important, fact about the discovery is that hardly anything about it was documented at the time. Six months would pass before Fleming published his results in 1929, and penicillin would remain at best a novelty, at worst a dead end, for another decade. By the time the magnitude of the discovery came to light, it’s certainly possible that the details had faded in Fleming’s memory, or that he recalled a sequence of events reminiscent of his earlier discovery without any intention to deceive. The debate about his reasoning (if any) and motives (ditto) continues, with no resolution in sight.
This wouldn’t be very important except for the significant matter of credit, which, as we shall see, would become a very thorny issue indeed. Insofar as any one person is associated with the discovery of penicillin—of antibiotics generally—in the public consciousness, it’s likely to be Alexander Fleming. It was considerably easier for Fleming to become the first hero of the antibiotic age if his great discovery were understood to be one he himself recognized immediately, rather than one about which he was mistaken—which was apparently the case, at least for a few months. The story that better fits the facts, one that was persuasively argued by the physiologist Robert Scott Root-Bernstein, is that Fleming hadn’t been doing a staph experiment at all; instead of testing a number of different pathogens, he was actually observing a number of different fungi. He was, therefore, surprised by the accidental introduction of the staph colonies, and not the other way around.
Which leaves unanswered the experiment’s goal, about which Fleming was famously unhelpful. Experiments need hypotheses, after all. If his goal was to observe staph colonies under different conditions, what were they? If, instead, his objective was something different, the problem vanishes. Root-Bernstein’s answer? Fleming was seeking a new source for lysozyme.
In 1928, lysozyme was still Fleming’s only notable discovery; even after he had become world famous, it remained so important to him that he felt obliged to observe, in his Nobel Lecture in 1945, that “Penicillin was not the firstantibiotic I happened to discover. In 1922, I described lysozyme, a powerful antibacterial ferment which had a most extraordinary lytic [i.e., destructive] effect on some bacteria.” And molds were an extremely promising source for antibacterial compounds like lysozyme. As early as 1876, John Tyndall described the effects of the mold: “On the 13th [December 13, 1875] a thick blanket of Penicillium” that had formed on meat “assumed a light brown colour, as if by a faint admixture of clay . . . the slime of dead or dormant Bacteria, the cause of their quiescence being the blanket of Penicillium. I found no active life in this tube, while all the others swarmed with Bacteria.” Joseph Lister himself had noted that bacteria grown in samples that included molds—formally, the one known as P. glaucum—failed to grow. It is suggestive that Fleming’s original notes show that he thought the original bacteria-inhibiting fungus was P. rubrum, not, as the mycologist Charles Thorn later showed, P. notatum. This is a lot easier to explain if Fleming was testing a number of different molds, in an ongoing search for something he knew existed—lysozyme—rather than something never before seen. He had been working diligently since 1922 examining mucus, sputum, blood, plasma, eggs, snail slime, flowers, and even root vegetables, looking for lysozyme, and the best explanation for the inconsistencies in Fleming’s years-later account is that he initially believed that he had found, in the Penicillium mold, not a new antibacterial compound, but an old one.
However dubious his later recollections of the discovery, Fleming deserves enormous credit for recognizing the potential of the compound he first called “mould juice” and, even more, for the experiments that followed. He and his assistant, Stuart Craddock, became penicillin farmers, cultivating crop after crop of Penicillium, and fertilizing them with protein from the heart of a bull. With sufficient quantities to begin testing, they established where the juice was effective—it killed not just staphylococci, but also streptococci, and a number of other bacteria—and where not—typhus, for example, was immune to it. And they established that it was harmless to nonbacterial cells. Though it would be more than thirty years before the mechanism was understood, the reasons for penicillin’s effectiveness against some, but not all, bacteria (and its lack of toxicity toward animal cells) were the same.
Since 1884, biologists had known that some varieties of bacteria stained a distinctive color when exposed to certain dyes, usually gentian violet. Such bacteria were named Gram-positive, for Hans Christian Gram, the Danish biologist who had discovered the phenomenon; those that didn’t take the stain were, predictably enough, Gram-negative.
When bacteria reproduce, they behave like a water balloon with a string tied around the middle: The original cell divides in half, while the cell wall is stretched and twisted. Penicillin (and related compounds) chemically weakens the cell walls of the bacteria as it splits—this is why penicillin only acts on bacteria when they’re dividing—but only for Gram-positive bacteria that are unprotected by the lipopolysaccharide outer membrane: staph and strep, but not typhus.* And not animal cells, which have membranes but not walls, and are therefore safe from the actions of penicillin.
Fleming and Craddock could demonstrate these facts empirically without knowing their underlying mechanisms (Fleming, in particular, was convinced that the compound was, like lysozyme, an enzyme). They could purify the “mould juice” and distill it, though not to the point that they were able to remove an unpredictable number of toxic impurities, nor did their fluids ever reach a concentration of more than 1 percent. They could even test it on a sinus infection afflicting Craddock. Unaccountably, though, Fleming never tried it on an infected animal. In the words of another researcher, “All Fleming had to do to demonstrate the curative effect of penicillin was to inject .5 ml of his culture fluid in to a 20 g mouse infected with a few streptococci or pneumococci. . . . He did not perform this obvious experiment for the simple reason that he did not think of it.”
One possible reason for the failure to test the fluid on a test animal was the difficulty of working with it; nothing Fleming and Craddock did could solve the problem of penicillin’s instability: Within days, sometimes hours, the stuff, evaporated now into syrup, would lose its effectiveness. As a result, though Fleming would return to penicillin research occasionally over the following three years, he actually spent more time during the 1930s on lysozyme. As late as 1940 he wrote of penicillin, “The trouble of making it” even as a local antiseptic “seemed not worthwhile.” He had run up against the wall separating bacteriology from biochemistry, and St. Mary’s had no one with the training needed to hurdle it. Craddock later recalled that “we knew very little when we began” using the classic technique of dissolving the compound in a solution of acetone (sometimes ether) and allowing it to evaporate. “We knew just a little more when we had finished.”*
St. Mary’s deficiency in well-educated chemists was both absolute and relative, especially as compared to Germany. Because German bacteriological research was largely taking place in industrial chemistry laboratories, the level of expertise in processes that occurred at the molecular level was almost unimaginably high. St. Mary’s Inoculation Department was probably Britain’s most sophisticated and successful bacteriological research facility; in May 1932, the Dean of St. Mary’s, Sir Charles Wilson (later Lord Moran) appealed to England’s greatest newspaper mogul, Lord Beaverbrook, asking for the £100,000 needed to build St. Mary’s into a world-class institution. Beaverbrook did come through with a generous contribution, around £60,000, but that was probably less than Klarer and Mietzsch spent just producing test compounds for Gerhard Domagk. Competing with I. G. Farben with nothing but donations from Britain’s aristocrats was a bit like a prep school taking the field against the New York Yankees.
The German dye conglomerates were even able to transform physicians into adequate chemists; in the case of Paul Ehrlich, brilliant ones. It was no accident that the argument at the core of Ehrlich’s 1908 Nobel Lecture supposed that the future of microbiology would be chemical rather than observational. For decades few outside Germany listened.
Or, if they did, they did not understand. Almroth Wright, despite his very impressive skills, was poorly equipped to appreciate the importance of chemical analysis and synthesis. Worse, Wright’s conviction that any successful attack on bacterial pathogens would emerge from the immune system rendered him uninterested in hiring competent chemists. In consequence, when Domagk’s sulfa drugs ignited a revolution in medicine in 1934, Fleming’s 1929 paper “On the Antibacterial Action of Cultures of a Penicillium” was all that existed to record his most important contribution to humanity’s war on infectious disease.
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At his death in March 1902, Cecil John Rhodes, the English-born founder of the British South Africa Company, was one of the wealthiest and most celebrated men in the world. He had built a hugely successful business empire—among other things, he founded De Beers Consolidated Mines, then and now the world’s largest diamond mining concern—alongside one of the more traditional sort: the eponymous colony, later independent country, he christened Rhodesia. However, since Rhodesia was renamed Zimbabwe* in 1980 by an administration that found his brand of colonialism more than a little offensive, the Rhodes name lives on today only in institutions founded after his death: Rhodes University in South Africa, and, of course, the Rhodes Scholarships.
The scholarship, which funds two or three years of study at one of the University of Oxford’s residential colleges, is awarded annually to college graduates from current and former British colonies, the United States, and—when world wars don’t intervene—Germany. As of this writing, there have been 7,600 Rhodes Scholars, including dozens of men and women who are even more celebrated than their benefactor: Nobel Prize winners, generals, a few professional athletes, cabinet members, senators, governors, Canadian and Australian prime ministers, and even one U.S. president. It’s a close call which of them has had the greatest impact on history, but there’s little doubt about the first Rhodes Scholar to change the world, a physician who departed Australia for the journey that brought him to Magdalen College in January 1922. His name was Howard Florey.
Florey was then twenty-three years old, the youngest of five children born in Adelaide to a first-generation Australian mother and an English émigré father. He had excelled at both St. Peter’s Collegiate School and at the University of Adelaide in every academic subject except math,* and at an exhausting number of athletic pursuits, including tennis, cricket, and football. His father’s death in 1918, while Florey was studying for his medical degree, freed him to apply for, and accept, the scholarship that brought him and sixty-one others to Oxford,* a freshly minted MD ready to begin his studies at the university’s Department of Pathology.
Pathology—broadly speaking, the study of the causes of disease—was, for obvious reasons, a young discipline in 1922, no older than the discoveries of Koch and Pasteur fifty years before. Oxford had started teaching it as a course in 1894, and achieved departmental status only in 1901. The same year Florey arrived in Oxford, the pathology department received a gift in the amount of £100,000 from the trustees of the estate of Sir William Dunn, a Scottish merchant banker and politician who had, like Cecil Rhodes himself, made a fortune in nineteenth-century South Africa. Though the planning and construction of the Sir William Dunn School of Pathology took four years—it wouldn’t open its doors until 1927—Florey’s timing could scarcely have been better.
In 1923, the young Australian earned First-Class Honours from the School of Physiology and the Francis Gotch Medal awarded to the department’s most promising researcher. In his notebook of that year, the secretary of the Rhodes Trust, Sir Francis Wylie, described Florey as “a first-rate man—ranks with our best.” Wylie wasn’t the only one to recognize his talent. One of Florey’s instructors, Sir Charles Sherrington, nominated him for the John Lucas Walker Studentship at Cambridge’s Gonville and Caius College in 1923, which paid a stipend of £300 a year—around £15,000, or $24,000, today—plus another £200 for equipment.
Florey’s second year in England was productive. He published four papers on a variety of topics, served as medical officer for an Arctic expedition, and, despite what even Florey himself recognized as a less than appealing personality—in 1923, he described himself in a letter to his future wife, Ethel Reed, as “developing into a rather nasty product”—even acquired friends; one of them, Charles Sutherland Elton, would become one of the founders of modern population ecology. He had found a mentor in Sherrington, who wasn’t just professor of physiology at Oxford, but, in 1923, president of the Royal Society, the world’s first and most distinguished scientific association, which meant that his endorsement was about as good as it got for an ambitious young researcher.*
During his third year he made his first connection with the Rockefeller Foundation.
Established in 1913 by the Rockefeller family “to promote the well-being of humanity throughout the world,” in 1923 the foundation was the world’s largest philanthropic enterprise, with a special interest in medicine and health. It had already established the world’s first School of Public Health at Johns Hopkins University, and a dozen more public health universities around the world, working to eradicate parasitical diseases like malaria and yellow fever.
There were a number of reasons behind the decision of the world’s wealthiest family to embark on a crusade to give away millions of dollars. One was surely to repair some serious image problems. After decades of building the Standard Oil trusts, and accusations of brutal competitive tactics in order to corner the entire oil industry, John D. Rockefeller, Sr., was not the most admired man in America. Well-meaning (and well-publicized) philanthropy could reverse some of this, but even there, the path was fraught. The behavioral and social sciences, including anything that touched on the nature of industrial relations, were strictly forbidden, especially after the 1914 Ludlow strike—better known as the Ludlow Massacre—in which dozens of miners and their families were killed at the Colorado mine owned by John D. Rockefeller, Jr. Pure science was the ticket: the purer, the better.
The really novel aspect of the foundation, though, was how the money was disbursed: Rockefeller, who had neither the time nor the inclination to decide on the foundation’s grantees (and in any case his involvement was widely regarded as the next thing to poisonous), wanted experts doing the selection. Scientists would recommend other scientists, the best and the brightest that could be found. One of the foundation’s executives, Wickliffe Rose, spent nearly a year in Europe asking the continent’s most prominent researchers for nominees that might deserve grants from the foundation’s International Education Board in order to—his words—“make the peaks higher” and let the results flow down the mountain to everyone else.* So-called circuit riders of the Rockefeller Foundation traveled the world, checkbook in hand, looking to identify the “future leaders in science,” and both Cambridge and Oxford were regular stops on their journeys.
In 1925, Howard Florey received his first Rockefeller Foundation fellowship, spending nine months, from September 1925 to May 1926, at various Rockefeller-funded labs in New York, Chicago, and Philadelphia, where he would establish a relationship with Alfred Newton Richards, the future president of the National Academy of Sciences.
Florey’s transatlantic connections would prove enormously important, as, too, would his links to continental Europe—he had spent part of 1922 and 1923 at labs in Copenhagen and Vienna—though forging them proved a challenge. By the 1920s, scientific research had become so much more international than it had been when Koch and Pasteur were dueling over the relative importance of vaccination and sanitation as to be unrecognizable. German, French, British, Swedish, and American scientists collaborated regularly, published together, and—this isn’t a contradiction—still fought fiercely over discoveries and priority. National affiliation wasn’t completely forgotten, but the real competition was between laboratories, not nations.
All that collaboration and competition, however, wasn’t free. Over the course of the next two decades, two different schemes for the funding and direction of scientific research were on offer. The industrial model, which had produced Paul Ehrlich’s Salvarsan, and would, soon enough, be responsible for Gerhard Domagk’s sulfa drugs, had the advantage of tremendous focus and discipline, but it was also highly dependent on confidentiality. The philanthropic model, on the other hand, benefited from collaboration and the ability to follow multiple lines of investigation, but even with the world’s wealthiest families supporting it, it was still underfinanced. Worse, unlike corporations like I. G. Farben, philanthropists, whether individuals like Sir William Dunn or trusts like the Carnegie and Rockefeller foundations, lacked ruthlessness. They were temperamentally ill-suited to disinvest from programs rapidly, and, as a corollary, foundation-led research had difficulty reinforcing success. Even with Lord Beaverbrook’s money, Alexander Fleming’s department at St. Mary’s couldn’t afford to hire the chemical talent needed to exploit the original penicillin discovery. The same story—brilliant and promising research in a constant scuffle for money—was about to play out at Oxford, with a somewhat different conclusion.
In 1926, Florey married Ethel, his former medical school classmate, now a doctor herself. Even by the standards Florey had established for bluntness and interpersonal tone deafness in his professional life, his courtship, conducted via slow-motion correspondence between England and Australia, had been stormy. It was calmness itself compared to the marriage that followed, though, which featured score settling of a particularly vitriolic (and difficult to read) nature. By the time the Floreys’ marriage was five years old, Ethel was complaining that her husband had sabotaged her career, while he, in turn, accused her of desertion, lack of affection, a disappointing frequency and variety in their sex lives, lousy cooking, and poor personal hygiene, even reminding his—deaf—wife that she was “not a physically normal woman.”
As miserable as Florey was in his marriage—or, to be accurate, as miserable as both Floreys were—it did little to divert his career. In 1927, he received his PhD from Gonville and Caius College at Cambridge, where he was appointed a lecturer in special pathology—a discipline about which he had known nothing before Sherrington took an interest in him, but which would be the subject to which he would devote the rest of his life.
Establishing himself in the field of pathology meant studying pathogens. For two years, Florey produced a huge volume of work, on subjects as varied as cerebral circulation, capillary action, and mucous secretions: an “experiment a day, including Sundays,” in the words of a colleague. He spent the summer of 1929 on another grant-paid trip to Madrid, mastering the art of cell staining with Professor Santiago Ramón y Cajal, the pioneering histologist and neurologist who had won his own Nobel Prize in 1906 for describing the cellular nature of the nervous system. And he began thinking about the gut. In 1931, Florey took his investigations to the University of Sheffield, which had offered him the Joseph Hunter Chair of Pathology. Four years later, he returned to Oxford.
By then, the Dunn School had been open for eight years, led by George Dreyer, a brilliant pathologist who had made his name analyzing diphtheria toxin. Specifically, and following in the steps of Paul Ehrlich, who had established the standard for measuring the toxin, Dreyer standardized the agglutinating power of blood serum into a single numerical value.
Standard setting is the sort of unexciting but vital scientific research that tends to be scanted in most histories. It was, however, hugely important, and about to become more so, as pathology and pharmacology became a matter of testing large numbers of different compounds: the more, the better. Consider the hundreds of azo-plus-sulfa side chains that were, at virtually the same time, being produced by Klarer and Mietzsch at Bayer’s tropical medical group. The ability to compare samples using a single number was critical, and Dreyer understood this better than anyone in Britain. Among his notable achievements while at the Dunn School was building Britain’s first Standards Laboratory, a repository for pure samples of many pathogens.
Dreyer died in 1934, which opened up what, in the ornate language of Oxford, was known as a “Statutory Professorship”—a position within the university that was, by tradition, administered by one of Oxford’s thirty-nine residential colleges.* For the Dunn School, that was Lincoln College, whose rector was importuned by a number of dignitaries supporting Florey’s candidacy. They included an old mentor—Charles Sherrington—and a new one: Edward Mellanby, the pharmacologist who had discovered vitamin D and demonstrated its relationship to the deficiency disease known as rickets. Mellanby had been one of Florey’s supporters on the Sheffield nominating committee, and had since become even more prominent as secretary of Britain’s Medical Research Council, which had been founded in 1913 as a publicly funded agency charged with financing medical research, initially on tuberculosis, but from 1920 on, on all diseases. In 1934, those funds were still extremely hard to come by, particularly as compared to the investments being made by the German chemical conglomerates, but the MRC did have one advantage that they lacked: a Royal Charter that authorized collaboration between researchers in academic settings like the Dunn and those in the chemical and pharmaceutical companies.
It was clearly the right moment for such collaboration, and the Dunn was the right place. It remained to be seen whether its new leader—Florey was named director in December 1935—had the right research agenda. It’s unclear when, or why, Florey started a series of investigations into the well-known but poorly understood phenomenon that the wall of the gastrointestinal tract was impermeable to bacteria—why potential pathogens didn’t infect the wall itself. A number of theories were currently popular, and one of them was the presence of the antibacterial compound lysozyme, discovered by Fleming eight years before.
Florey had been interested in lysozyme ever since, and had even gotten a grant to extract it in a pure form from egg whites. The need for purifying lysozyme led directly to one of the most important decisions in the history of medical research, a decision not about technique or theory, but personnel. Florey, like Domagk, needed chemists to give him the raw material (or, more precisely, the refined material) for his experiments. Even while at Sheffield, he had been begging the Medical Research Council for just such a collaborator, to purify Fleming’s lysozyme and identify its substrate: the molecular component on which the compound did its work. At Oxford, though, he had considerably more to offer, and, in 1936, he finally got his chemist: E. A. H. Richards, from one of the world’s most innovative organic chemistry departments, Oxford’s Dyson Perrins Laboratory.* By 1937, Richards had succeeded where Fleming had failed, producing lysozyme in pure form. Even more consequentially, in the same year Florey charged another staff chemist with the job of identifying lysozyme’s substrate. The staffer was a Jewish émigré from Germany named Ernst Chain.
Certainly, Chain’s background seems, on first glance, to have been wildly different from Florey’s. Born in Berlin in 1906, the son of a German mother and a father who had emigrated from Russia’s “Pale of Settlement”—the region in eastern Russia that, from 1791 to 1917, had permitted permanent Jewish residency—to Germany, where he studied chemistry, changed his name, and opened the Chemische Fabrik Johannisthal Adershof, a factory that produced pure elements like copper and nickel for industrial use.
This hides more than a few similarities. Florey’s father, too, was an immigrant, though from England to Australia. Both fathers took the entrepreneurial path followed by immigrants everywhere, in John Florey’s case starting a business manufacturing boots. Both fathers died while their sons were still at school, Florey’s in 1918, Chain’s a year later, a financial hardship for both families. The death of Chain’s father left his family—including Ernst’s mother and sister, Hedwig—somewhat strapped, though not so distressed that Ernst could not take advantage of Germany’s extraordinary educational system; and in 1927, he graduated from Friedrich-Wilhelm University (now Humboldt University). In 1930, he added a D.Phil. from the Institute of Pathology at Berlin’s Charité Hospital, the alma mater of more than half of Germany’s Nobel Prize winners, including Emil von Behring, Robert Koch, and Paul Ehrlich. In April, with his new degree and, as he later recalled, £10 in his pocket, he left for England, leaving his mother and sister behind.
Within two years, Chain was publishing papers in scholarly journals and, despite a passport endorsement that should have forbidden him from accepting “paid or unpaid employment,” had joined the chemical pathology lab at University College Hospital. In May 1933, despite that pesky business about employment—he was then living on a stipend of £250 annually from the London Jewish Refugees Committee and Liberal Jewish Synagogue—he got a job in the Department of Biochemistry at Cambridge, working for his own mentor: Frederick Gowland Hopkins, who had been awarded half of the 1929 Nobel Prize in Physiology or Medicine (for the discovery of vitamins) and had been, since 1930, president of the Royal Society.*
And, like Florey, Chain was ambitious, confident, and resourceful. In the words of one biographer, he provided an “inexhaustible flow of ideas and suggestions for overcoming difficulties,” which would have made him a desirable colleague but for an unhealthy dose of arrogance and a habit of condescending to others, aspects of his personality he was either unable or unwilling to hide. Unsurprisingly, this was, again as with Florey, the cause of frequent irritation among his coworkers. They put up with him, and even sought him out, for his excellent experimental technique, and for a truly remarkable, near-photographic memory. Chain’s ability to summon every pertinent reference in the scholarly literature of biology and chemistry without recourse to a library made him the 1930s equivalent of a laboratory connection to the Internet. A dozen different colleagues would later recall his ability not merely to quote page and volume from relevant journal articles, but to quote text verbatim, even citing where on the page the important passage would be found.
Chain had another advantage in the lab. Long before he had become a scientist, he was a piano prodigy, good enough to give concerts in Berlin in his teens, and one who maintained his technique with constant practice. He was gifted enough that at least one account has him visiting Buenos Aires on a concert tour in 1930; certainly, as late as 1933 he was so torn between biochemistry and music that he interviewed for a job in the BBC’s orchestra.
Chain’s musicianship gets mentioned frequently in biographies, often as evidence of the “artistic temperament” that made him a creative experimentalist. Far more valuable, though, for the laboratories of the day, was Chain’s combination of a pianist’s muscle control and hand-eye coordination. Steady hands are as valuable to a chemist as they are to a stage magician; the simple experimental technique of titration depends on adding one liquid to another, a droplet at a time, in order to observe the beginning and end of a chemical reaction in a precisely calibrated vessel. Growing crystals by diffusing one solvent into another has to be done slowly and steadily enough to keep the boundaries between them distinct. Small wonder that Chain’s colleagues were as likely to recall his deftness with beaker and pipette as they were his memory.
Credit: Wellcome Library, London
The Dunn School team: Howard Florey (back row, second from left) and Ernst Chain (back row, second from right)
Florey didn’t have Chain in mind when he made recruiting biochemists a priority for the Dunn after he took over in 1935. His first choice for the job was another of Frederick Gowland Hopkins’s Cambridge protégés, Norman Pirie, a Scottish biochemist and virologist. Pirie was either uninterested or unavailable, and it was Hopkins who suggested Chain. “I find his biochemical knowledge is more than merely adequate . . . he has really become a well-qualified biochemist. . . .”* Chain was interested in the job. As he would later write, his “principal motivating principle . . . was always to look for an interesting biological phenomenon which could be explained on a chemical or biochemical basis, and attempting to isolate the active substances responsible for the phenomenon and/or studying their mode of action.”
Of course, he wrote that long after he had departed Oxford as one of the most famous scientists in the world. When he arrived there, his priority was less on the disinterested search for scientific explanation and more on, well, the toys. After training in the state-of-the-art facilities at Berlin’s Charité Hospital, he had been regularly, and loudly, disappointed at the quality of the equipment available at Hopkins’s lab in Cambridge. So, when he arrived at the Dunn, and Florey’s chief technician, Jim Kent, escorted him to his lab and Chain saw—through the window of the neighboring Dyson Perrins lab—a Soxhlet extractor (a fairly rare piece of lab apparatus designed to extract lipids from solids, and used for all sorts of purifying exercises), his eyes grew as large as a child’s visiting a chocolate factory. Asked by Chain if the Dunn School used them, Kent said he believed they had one, to which Chain said, “One! I shall want ten!”
If only. The Dunn was well equipped only by the standards of other British laboratories, and Florey’s group still depended on the unpredictable largesse of individuals and foundations, and modest subsidies from the Medical Research Council. In the middle of a worldwide depression, none of them was what one might call generous. When Chain arranged for enlarging and modernizing the lab’s refrigerator—it had been manually operated by a steward, who turned on the compressor when he thought it was getting too warm—and exceeded the budget by £15, it “caused a terrific upheaval, and Florey never forgot this incident and reminded me of it until I left the Institute.”
Over the course of the next year, Chain worked diligently (and parsimoniously) on everything from snake antivenin to protein proteolysis, despite some disabling but undiagnosed diseases that caused him bouts of such severe depression and anxiety that he began keeping a health diary, where, in August 1935, he carefully noted his “periodic fear attacks.” Part of his anxiety was, no doubt, financial; with the title of departmental demonstrator, he was paid only £200 annually, and frequently enough needed to get advances on even this modest sum, which he supplemented with the occasional grant.
By the beginning of 1936, Chain had started another research program, this time on skin cancer, and needed an apparatus for measuring the very small amounts of oxygen uptake during the metabolism of cancerous tissues. While devices, known as respirators, existed for less granular investigations, Chain needed something on an order of magnitude more sensitive. To design his “microrespirometer,” he suggested one of his former Cambridge colleagues to Florey. Florey agreed, but—surprise of surprises—while he could afford the man, he had no money for the needed equipment.
Once again, the Rockefeller Foundation was tapped. Florey wrote to Daniel P. O’Brien, a former circuit rider, now associate director of the foundation’s Division of Medical Sciences, begging for money to expand “the chemical aspect of Pathology.” Soon after, O’Brien’s boss, Wilber Tisdale, visited Oxford, agreed with the request, and approved a grant that allowed Florey to buy “balances, micro balances, vacuum distillation apparatus, etc. to a value of £250.” Far more important than the equipment, though, Florey also agreed to hire Chain’s onetime colleague, a mechanical genius named Norman Heatley.
Heatley was then barely twenty-five years old, a newly minted PhD from Cambridge, and, luckily for his colleagues, possessed of a very different temperament than both Florey and Chain. Heatley even looked different. Chain was mustached, short, and intense, his head always set slightly forward, as if ready to attack; Florey was square jawed and robust, laconic and brusque. Heatley was tall, elegant, and slender. Where Florey and Chain were confident, verging on—generally passing over into—arrogant, Heatley was diffident, perhaps to a fault: He was known to have said, “I was a third-rate scientist whose only merit was to be in the right place at the right time.” He was also unfailingly courteous, so much so that he was regularly shocked by the egotism and ambition regularly on display at the Dunn. Certainly he got on better with Florey than with his putative supervisor, Chain, especially after the latter insisted on a credit in the journal article that described the microrespirometer, despite the fact that, as Heatley recalled four decades later, the device “was wholly my conception and design” and that Chain had demanded credit out of ambitious careerism.
The disputed article, which would eventually appear under the title “A New Type of Microrespirometer” in the Journal of Biochemistry in January 1939, isn’t a terribly important scientific paper, but it is a gold mine of foreshadowing. First, it revealed Florey’s special talent for preserving the morale, and so the productivity, of all his research assets; when Chain claimed that he deserved credit as an author for the paper, Florey was able to give it to him without simultaneously angering Heatley (at least, not very much). In the final article, as directed by Florey, Heatley is the first-named author, but Chain is the last—in the world of scholarly publishing, the second-best spot. (In between Heatley and Chain was a research fellow at the Dunn named Isaac Berenblum, who would become a world-famous oncologist after emigrating to the new state of Israel in 1949.)
More foreshadowing: On display are not only Florey’s careful management style and Chain’s ambition, but also Heatley’s great talent for building lab equipment out of spare parts and discards. Describing the magnetized iron balls needed to mix the droplets under investigation, he wrote, “Steel-bearing balls, 1/16 in. in diameter, are given several coats of Bakelite varnish . . . the balls are then heated to 100° in paraffin wax for some minutes, the surplus wax being removed by rolling the balls on hot filter paper. . . . They are then rolled in the palm of a warm, but clean and dry hand with some well washed kaolin. . . .”
The article was the first time the peculiar mix of talents of the Dunn team stood revealed. It also marked, or rather caused, a permanent breach in the relationship between Heatley and Chain. From that moment forward, at Heatley’s insistence, and with Florey’s tacit approval, all communication and direction for the young man from Kent would come from the Dunn School’s director, rather than its chief biochemist.
By this time, Chain had plenty of other subjects to keep him busy, most especially the one that had been Florey’s reason for bringing him—and E. A. H. Richards—to the Dunn in the first place: finding the substrate for lysozyme. In 1937, assisted by one of that year’s Rhodes Scholars, a medical student from Missouri named Leslie Epstein, Chain had found his answer: The substrate was determined to be a polysaccharide, which meant that lysozyme was a polysaccharidase, whose (mild) antibacterial action was that it broke down the polysaccharides (some of them, anyway—E. coli alone makes more than two hundred different polysaccharides) that coat the cell walls of bacteria. Epstein had found the subject for his thesis: “The Actions of Certain Bacteriolytic Principles.”
Around the same time, Florey found Alexander Fleming’s 1929 paper on penicillin.
No one knows precisely how the paper came to the attention of the Dunn investigators, or even which of them first read it. To his death, Chain was adamant that Florey never thought about penicillin until Chain suggested it. “Something seemed to click in my [i.e., Chain’s] mind” after reading Fleming’s paper. Florey was equally insistent that he had brought it up to Chain. All that can be known for certain is that, during the preceding eight years, virtually no other researcher had cited Fleming’s work.
Fleming’s discovery was a classic dead end: an interesting compound that was so unstable that even its discoverer couldn’t reliably produce it for future experiment, nor could anyone else. Though Florey’s predecessor at the Dunn, George Dreyer, had been intrigued enough by Fleming’s mold to secure some for the pathology lab, he had done so for dozens of potentially interesting compounds, and no one had been any more fortunate than Fleming himself in understanding its actions. No one, that is, until 1937, when Florey and Chain began planning an ambitious survey of all the antibacterial substances produced by microorganisms. The planned survey would include dozens of different strains of bacteria, but also fungi, particularly the Penicillium molds.
The first experiments of the new survey, though, were still focused on lysozyme and other potential antibacterial substances, since penicillin, in its decidedly impure, “natural” form, was such an unreliable antibacterial agent. In any case, Chain believed penicillin to be a kind of “mould lysozyme,” an enzyme that acted, like Fleming’s egg-white lysozyme, on bacterial cell walls, but also on pathogens like staph and strep, so research on lysozyme was likely to be applicable to penicillin anyway.
In one well-remembered discussion over afternoon tea, Florey reminded his listeners that penicillin was also notoriously difficult to work with; not only had Fleming been unable to stabilize his own compound, Harold Raistrick, a skilled and experienced biochemist, had no better luck. Chain reacted by saying Raistrick couldn’t be a very good chemist, since it “must” be possible to produce it in a stable form. Whether deliberately or not, Florey had challenged Chain.
Chain, who was as competitive as a pit bull, responded, though it is worth a reminder that both he and Florey saw the research as an interesting scientific challenge far more than as a way to add to medicine’s therapeutic arsenal. Prontosil and the other sulfanilamides were rightly regarded as revolutionary therapies, and there seemed little need or desire to supplant them.
Chain was already investigating the substance produced by Pseudomonas pyocyanea (the bacterium responsible for, among other things, septic shock and a number of skin infections; since the 1880s, extracts of P. pyocyanea had been shown to destroy other bacteria . . . and to be highly toxic to mammals), and the somewhat more promising Bacillus subtilis, a hardy microbe with some demonstrated ability to stimulate the immune system. To them, he added the remaining frozen samples of Penicillium notatum that Dreyer had left behind, but his first results were unimpressive. He could study the mold if he had enough, but it was “impracticable to grow the [Penicillium] mould and carry out chemical studies simultaneously.”
Heatley stepped into the breach. Despite his self-effacing modesty, he was, in the words of Gwyn Macfarlane, a hematologist at the Radcliffe Infirmary and later one of Florey’s biographers, “a most versatile, ingenious, and skilled laboratory engineer on any scale, large or minute. To his training in biology and biochemistry, he could add the technical skills of optics, glass and metalworking, plumbing, carpentry, and as much electrical work as was needed.” Most important of all: “He could improvise—making use of the most unlikely bits of laboratory or household equipment to do the job with the least possible waste of time.”
When he was drafted to increase the yield of the antibacterial substance produced by the Penicillium mold, he knew relatively little about it, except that the fungus grew adequately on agar, but did best in shallow vessels, no more than 1.5 centimeters deep. At that depth, the branchlike mycelia of the mold could grow above the surface of the agar, and then dry out. Once dry, yellow drops of Fleming’s “juice” formed on the dried-out mycelia and could be collected using a glass pipette. Even more valuably, other penicillin droplets settled into the agar itself and turned it yellow. The most productive time for penicillin “farming,” therefore, was just after the broth turned rich enough to be harvested, but before it became so saturated that the agar couldn’t grow another batch. By careful observation, Heatley learned to identify the agar’s phase of maximum productivity.
Agar offered a good base for growing Penicillium, but continued to produce frustratingly small amounts of Fleming’s broth. The stakes were high; only with significant quantities of broth could any investigation proceed, and Heatley knew it. He fertilized it with everything he could find on the Dunn’s shelves: nitrates, salts, sugars, glycerol, meat extracts. He dosed the media with oxygen and CO2. In December 1939, he tried adding brewer’s yeast, which improved the yield only slightly, but did cut the time it took for the mold to produce the broth from three weeks to ten days.
It took months before Heatley determined the best recipe. First, he incubated the fungus on a nutrient solution known as Czapek-Dox medium: a stew of inorganic salts, sugar, and agar. Once the mold bloomed, brewer’s yeast was added. In days, a film formed on top of the medium, and soon thereafter, green spores of the Penicillium would appear. Over the course of ten days, the fungus would grow, after which Heatley drew off the penicillin-laced broth and replaced the growth medium, twelve times if he was lucky, two or three times if not.
This provided sufficient amounts of broth, but did little to gauge its strength. Though Fleming and those who followed had been able to demonstrate the antibacterial activity of the liquid extracted from mold, what they had wasn’t penicillin, but a broth of which penicillin was a component. How much of the liquid was biologically active? No one knew. Heatley needed a yardstick for measuring antibacterial activity, and once again he found an ingenious solution: He cut disks out of the bottoms of Petri dishes and replaced them with glass tubes, creating concave indentations in the center of the vessels, into which colonies of bacteria were introduced. He then added measured amounts of the cultivated yellow broth, and noted the size of the bacteria-free halo around each cylinder. The bigger the halo, the more potent the compound.
If Heatley was legendarily resourceful in making equipment for pennies, Florey was no less so in collecting the pennies themselves, wherever they could be found. The use of the word “pennies” is not accidental; to call the Dunn experimental program impoverished is to flatter. At the 1938 Physiological Congress in Zurich, Florey buttonholed Edward Mellanby to beg the Medical Research Council for what seems, in retrospect, a ridiculously tiny sum: £600. Even when he got it, it was scarcely enough. At one point, Florey told Chain that the lab had completely exhausted its funds, and that he must stop ordering everything, up to and including glassware. Though the Medical Research Council finally agreed to renew a portion of the lab’s grant for 1939—for Chain, £300 a year for each of the next four years, plus an additional grant for expenses of £250 annually through 1940—it kept it just north of starvation. When Florey learned that Oxford was proposing to cut the operating budget they supplied to the Dunn because a new university heating plant would lower the laboratory’s utility costs, Florey wrote back, “I have struggled to keep the place warm on money I ought to have devoted to research.”
Matters nearly came to a head in the summer of 1939, when the grant from the Medical Research Council that paid for Chain’s research on the penicillin project was about to expire. Florey might not have loved Chain’s company, but he recognized his value and was determined to find the money needed to keep his biochemist fed, housed, and not too surly for work. Their grant application not only identified fungi as a promising source for antibacterial compounds, but announced the status of their experiments on Alexander Fleming’s nearly forgotten eleven-year-old discovery.* For the first time, penicillin was an explicit part of their research program.
Florey was taking no chances. He sent a virtually identical grant proposal off to the newly established Nuffield Provincial Hospitals Trust (another charity built on a wealthy man’s will, this time a bequest from William Morris, who had made his fortune selling Morris Garages sports cars—MGs), and another to the usual suspects at the Rockefeller Foundation, though by a circuitous route. The original request was sent to the Rockefeller offices in France on November 20, and was then forwarded to the New York office ten days later, where the director of scientific research, Warren Weaver, wrote back, “The application of Florey appeals to me, but I seriously question whether a three-year grant is justified under present circumstances. . . .”
The “present circumstances,” were, of course, the Second World War, which had commenced with Germany’s invasion of Poland on September 1, and within days, declarations of war from France and Britain. By the time Weaver received the Dunn School’s grant application, Poland had been divided between Germany and the Soviet Union, the United States had passed the Neutrality Acts (which allowed France and the United Kingdom to buy arms), and the estuaries of the Thames had been mined by U-boats. The threat of war had already had an impact at the Dunn; Chain, as a refugee perhaps more fearful than most of a German invasion, and more grateful to Britain for offering sanctuary, had volunteered for a Red Cross Certificate in First Aid, and, after becoming a British subject in April 1939, joined the Oxford City Council Air Raid Precautions Department. Heatley was unable to leave England for a fellowship in Copenhagen, and—at Florey’s behest—stayed in Oxford.
Initially, the transition to open hostilities would shave budgets for research, particularly of the “interesting scientific challenge” sort. After some back-and-forth, including reassurances from Weaver that the grant would be renewed as long as the Dunn team showed progress, the money started flowing again, though, as always, through a very narrow straw. Though Florey’s original request to the Medical Research Council for studying penicillin as a therapeutic agent in vivo was for a mere £100, the MRC actually came through with only £25. Luckily, on February 19, 1940, the Dunn team learned that the Rockefeller grant had been approved, with the first payment scheduled to arrive March 1.
Tiny budgets notwithstanding, the Dunn research was progressing, and progressing rapidly. By March 1940, Heatley’s methods had improved yields so much that instead of providing Chain with a milligram of broth at a time, he could make a hundred times as much.
However, the trick of extracting penicillin from the broth in a stable form, which had eluded Raistrick and others, was still undiscovered. Fleming had tried to separate the active ingredient in his mold juice using a simple chemical technique: Dissolve the stuff in ether, which would evaporate quickly, leaving behind concentrated penicillin. It had failed almost completely. Harold Raistrick, a far more skilled chemist, used a method of separation known as liquid-liquid extraction: Pour the mixture into a “separatory funnel,” a piece of apparatus that looks like an inverted teardrop, with a funnel at the top, and a stopcock at the bottom, with a flask underneath. In the top chamber, liquids separate into different layers based on densities, with the heavier “aqueous” layer, containing ions, or charged particles, at the bottom, and a neutral, uncharged “organic” layer at the top. Shake vigorously, open the stopcock, let the bottom layer flow out, and you have your extract.
The key, then, was to add a charge to the molecules that composed the desired portion of the broth.* There are a couple of ways of charging a neutral substance, but one of the most effective is to acidify it, since what makes acids acidic is a freely given positively charged hydrogen atom: a proton. Give a proton to a neutral compound, and it becomes charged. The greater the charge, the greater the amount of the previously neutral substance that heads to the bottom of the separatory funnel. This was Raistrick’s strategy. By adding acidified ether to the solution, a little at a time, he was able to separate the mold juice from superfluous fluid, leaving behind a concentrate of about one-fifth the volume with which he began.
However, when he evaporated the ether (easy enough; it’s a very volatile substance) expecting to find a concentrated form of penicillin in the residue, it had completely vanished: None of the antibacterial activity that Fleming had identified in the mold juice remained.
In the intervening years, no one had been able to improve on Raistrick’s method. This meant that, despite Heatley’s resourcefulness in cultivating Penicillium’s precious broth, no one knew how to convert his harvest of broth into a stable source of penicillin.
Enter Heatley, again. He first tried to stabilize the compound at different temperatures and pH levels, slowly adding alkali, or base, salts to the mixture to get it back to neutral, but it wasn’t a very practical method, the equivalent of baking a soufflé over a campfire. His second idea, what he later called “laughably simple,” reextracted the compound from the acidic ether solution into a neutral medium—water—by taking another trip through the separatory funnel, and then gave it another charge, this time by exposure to a base. On March 19, 1940, he did just that, filtering the mold broth through parachute silk (in order to remove solid particles), then mixing it with ether, which caused it to layer and separate. The ether-plus-penicillin mixture was then mixed with alkaline salts, dissolved in water, and separated again; only this time, the penicillin was in the aqueous layer. And, unlike the ether-plus-penicillin mixture that had lost all its potency for Raistrick, the new mix was stable, even after eleven days at room temperature. A source for experimental quantities of far purer penicillin—though the term is used loosely; while the concentration of penicillin in Fleming’s broth was only one part per million, Heatley’s first batch was still no better than .02 percent pure—had been found.
And Chain was ready to experiment with it. Almost as soon as he received the purer extract of penicillin, he directed John M. Barnes, one of the few researchers at the Dunn licensed by the Home Office to perform animal tests, to inject the entire stock of penicillin extract, 80 milligrams, into the abdomens of two mice.
It isn’t clear what he expected in the way of a reaction. Over the preceding year, Chain’s supposition about the nature of penicillin—that it was a complex protein molecule, probably an enzyme, like lysozyme—had been fading. Though the explicit aim of the 1939 grant was the “preparation from certain bacteria and fungi of powerful bactericidal enzymes, effective against staphylococci, pneumococci, and streptococci,” by the beginning of 1940, Chain had completed a number of experiments that proved penicillin couldn’t be a “bactericidal enzyme,” or indeed a protein of any sort. In one experiment, penicillin dialyzed—broke into component parts—when forced through cellophane tubes—which is something that proteins, because of their size, don’t do; in Chain’s words, his “beautiful working hypothesis dissolved into thin air.” If penicillin had been a protein, the mice would have exhibited an immune response: swelling, perhaps, or inflammation. But they didn’t. The mice might as well have been given saline solution; there was no impact. Penicillin definitely wasn’t any sort of protein.
The bad news was that Chain hadn’t found a complex molecule, but a relatively simple one, though, again in Chain’s words, “it became very interesting to find out which structural features were responsible for the instability. It was clear that we were dealing with a chemically very unusual substance.” The good news was that the mice tolerated it almost completely, which suggested that, unlike almost every other promising antibacterial compound, it seemed safe. Even better: the extract, once excreted in urine, was a) nearly as brown as the compound injected, and b) still strongly bactericidal.
The Dunn team was on the verge of isolating a substance that killed pathogens without damaging their hosts. Which made it highly interesting medically.
Howard Florey made penicillin the Dunn School’s number one priority. Two months after Chain’s first test, at 11:00 A.M. on May 25, 1940, Florey infected eight mice with Streptococcus pyogenes, the pathogen responsible for infections like strep throat, impetigo, erysipelas, and even the flesh-eating nightmare, necrotizing fasciitis. At noon, two mice were given 10 milligrams of penicillin, and another two were given 5 milligrams. Follow-up doses were given at 4:15, 6:20, and 10:00. Just before midnight, as Heatley recorded in his lab notebook, “one mouse got up and staggered about for a few seconds, then fell down, twitched once or twice, and was dead.” By 4:00 A.M. on May 26, all four of the controls had died.
All four of the treated mice survived.
The following day, as the Battle of France raged, nearly seven hundred British ships and small craft started evacuating what Prime Minister Winston Churchill called the “root and core and brain of the British Army” from Dunkirk. It would be four years before they would return, on June 6, 1944: D-day. When they began the liberation of France, the most important and valuable medicine used by their medics and field hospitals would be the same substance that had saved four of the Dunn School’s mice.