Miracle Cure: The Creation of Antibiotics and the Birth of Modern Medicine 1st Edition

FOUR

“The People’s Department”

The Dunkirk rescue was the best news that Britain would experience for months, as the seemingly unstoppable Wehrmacht conquered Belgium, Norway, the Channel Islands, and—on June 25—France. The Dunn team was acutely aware of the German threat, but even more focused on the progress of their own investigations. The success of the May 25 experiment launched the already highly energetic Howard Florey, and everyone else at Dunn, into another gear. Within days, more successful experiments with mice followed, revealing the potency of even highly diluted concentrations of penicillin.

It also exposed a pressing need for more staff. Though the mice experiments showed the effectiveness of the penicillin broth, they revealed little about why it worked. Bacteriologists Duncan Gardner and Jean Orr-Ewing were brought on board to investigate the mechanism by which the compound was performing its magic. Florey himself, along with Jim Kent, planned and executed a series of experiments comparing the results of different dosages of both streptococci and penicillin at different stages of infection.

The chemical investigations were accelerating. The Dunn team needed more penicillin at more powerful concentrations, and that could only happen with a better understanding—any understanding, really—of penicillin’s chemical structure. A twenty-seven-year-old chemist named Edward Penley Abraham was assigned to work with Chain on the daunting problem of purification.

Abraham and Chain were a well-matched team. They used a variety of purification techniques aimed at improving the compound’s potency, as measured by the ratio of the active ingredient to the rest of the solution: then as little as 0.5 milligrams per liter, less than one part in a million. The most effective technique turned out to be freeze-drying: First, dissolving the penicillin with dry methanol, diluting it with water, then freezing the liquid. Second, exposing it to a high vacuum to sublime the solvent, transforming it directly from a solid to a gas. Third, after all the sublimable solvent has been removed, separating the rest of it by exposure to high heat. The real eye-opener for the two chemists was the discovery that, even when diluted to one part in a million, penicillin still stopped bacteria from growing, which made it at least twenty times more powerful than the strongest sulfa drugs.

This was promising and frustrating in equal measure. Penicillin might be the most powerful and effective medicine ever discovered, but it was also one of the most difficult to produce. The lack of raw material was the most daunting bottleneck for further research. How, then, to produce more? Despite the grants from the Rockefeller Foundation and the Medical Research Council, Florey was still scrambling for money, and now needed larger-scale production resources.

One intriguing possibility would be to enlist Britain’s pharmaceutical industry, whose companies, though generally far smaller than Germany’s huge conglomerates, nonetheless possessed manufacturing expertise, investment capital, and, especially, a powerful interest in any compound with the potential to treat infectious disease. Glaxo, founded in New Zealand in 1880, had expanded into the United Kingdom less as a pharmaceutical company than as a manufacturer and processor of milk fortified with vitamin D;* by the 1930s, they were the country’s largest producer of nutrition products. Beecham’s Pills were Britain’s most popular laxative, and the foundation of Beecham Limited. Imperial Chemical Industries, the result of an I. G. Farben–style merger between competing companies (including Nobel Explosives and the British Dyestuffs Corporation), had been, since 1926, by far Britain’s largest chemical company, but, unlike the Germans, had little or no interest in pharmaceuticals. The small chemical manufacturing firm Kemball, Bishop and Company showed some interest, visiting the Dunn in March 1940 accompanied by Sir Henry Dale, president of the Royal Society, but its resources weren’t up to the task.

More promising was Burroughs Wellcome, then Britain’s most technologically sophisticated pharmaceutical company. The company had been founded by two American expatriates, Henry S. Wellcome and Silas M. Burroughs, graduates of the Philadelphia College of Pharmacy, who had decamped to London and opened for business in 1880. Sixteen years later, the forward-thinking Americans had built Britain’s first industrial biological research facility, the Physiological Research Laboratories, largely to produce the company’s own version of Robert Koch’s antidiphtheria serum.* When two chemists from Burroughs Wellcome visited the Dunn in July 1940, they politely declined Florey’s suggestion that they take on the challenge of purifying and producing penicillin.

With no other takers, the Dunn would have to produce the penicillin it needed for research in-house, making up in ingenuity what was lacking in money. What this meant, in practice, was throwing the job to Norman Heatley.

Heatley’s first challenge in producing sufficient quantities of the precious substance was a shortage of properly sized vessels for growing the mold itself. Faced with extraordinary difficulty in procuring appropriate glassware (or anything; Florey was so penurious that he had the Dunn’s elevator shut down in order to save £25 annually), Heatley turned to larceny. Pie trays and baking dishes mysteriously vanished from the Dunn’s kitchens. Sixteen bedpans likewise disappeared from the Radcliffe Infirmary, to reappear in the pathology department’s labs.

Over the course of 1940, Heatley continued to improve his technique for extracting penicillin from the broth produced in his bedpan factories. The most effective process he found depended on a truly remarkable machine, able to mix ether with penicillin, acidify it, and separate the concentrated broth.

 L0032175 PENICILLIN: apparatus used in the production Credit: Wellcome Library, London. Wellcome Images images@wellcome.ac.uk http://wellcomeimages.org Apparatus for producing penicillin. Norman Heatley collection. PP/NHE/A/2/1/5 Published:  -   Copyrighted work available under Creative Commons Attribution only licence CC BY 4.0 http://creativecommons.org/licenses/by/4.0/

Credit: Wellcome Library, London

The modified ceramic bedpans, used by the Dunn School team to grow penicillin broth

The homemade apparatus in operation resembled nothing so much as a Rube Goldberg cartoon.

1.   Three bottles—of broth, ether, and acid—are held upside down in a frame, until

2.   The glass ball stopper in the bottle containing broth is moved aside; liquid flows

3.   Into a glass coil surrounded by ice. Once cooled, the acidified liquid combines with acid from bottle number three and is jet-sprayed in droplets

4.   That arrive in one of six parallel separation tubes. Meanwhile

5.   The stopper on bottle number two, containing ether, is moved aside, releasing ether into the bottom of the whole arrangement. The filtrate in the separation tube is sprayed into a tube of ether rising in a four-foot-long tube. As penicillin has a chemical affinity for the ether, it transfers into that tube, leaving the remaining components of the original broth behind, to be drained out.

6.   Then, the penicillin-plus-ether (later acetate) solution is introduced into another tube, with slightly alkaline water. The penicillin-plus-water mixture—about 20 percent of the volume of the filtered broth that started the whole rigamarole—was drawn off.

Remarkably, Heatley’s collection of discarded junk and baling wire—the hole in the glass needed to produce the right-sized droplets was made by Heatley pushing the point of a sewing needle through hot glass; a cast-off doorbell rang when each bottle was filled—could turn about twelve liters of broth into two liters of decidedly impure penicillin in an hour. A quantity of penicillin sufficient for experimentation (on mice, at any rate) had been guaranteed.

Credit: Science & Society Picture Library

The filtration machine built by Norman Heatley to purify penicillin

Florey decided it was time to go public. The Lancet of August 24, 1940, contained half a dozen articles. Two of them—topically enough, given the Blitz—were on treating blast injuries to the lung. Another article was on meningitis; one described the orthopedic trauma known as “locking wrist.” Right in the middle, though, was the world changer: “Penicillin as a Chemotherapeutic Agent,” by E. Chain, H. W. Florey, A. D. Gardner, N. G. Heatley, M. A. Jennings, J. Orr-Ewing, and A. G. Sanders. (Florey, having already suffered through the sniping between Chain and Heatley over authorial credit, deferred to the alphabet.) The first line of the article reads, “In recent yearsinterest in chemotherapeutic effects has been almost exclusively focused on the sulphonamides and their derivatives. There are, however, other possibilities. . . .”

The possibilities were detailed in the following two pages, including results from the five key studies performed at the Dunn since March, in which as many as seventy-five mice had been exposed to pathogens as varied as staphylococcusstreptococcus, and clostridium: “During the last year methods have been devised here for obtaining a considerable yield of penicillin, and for rapid assay of its inhibitory power. From the culture medium a brown powder has been obtained which is freely soluble in water. It and its solution are stable for a considerable time and though it is not a pure substance, its antibacterial activity is very great. . . .”

Eleven years and five months after Alexander Fleming had published “On the Antibacterial Actions of Cultures of a Penicillium,” his discovery had finally been revealed as more than just a Petri dish curiosity: “The results are clear cut, and show that penicillin is active in vivo against at least three of the organisms inhibited in vitro.

Even before the Lancet article appeared in August, work at the Dunn had been proceeding on three intersecting tracks. Norman Heatley and the technical staff continued to improve the processes by which penicillin-rich broth was grown, and from which the active ingredient could be extracted. The biochemical team, primarily Chain and E. P. Abraham, were subjecting the compound to a series of experiments intended to establish its structure. And the bacteriologists and pharmacologists were designing new ways to test the antibacterial properties of the compound on laboratory animals.

It is a testimony to Florey’s great gifts as a scientific administrator that the projects—production, analysis, and effectiveness—all succeeded brilliantly, in spite of the enormous amount of friction produced by the Dunn’s collection of strong personalities. The achievement is notable even though it might be said that at least some of Florey’s challenges in managing the lab were self-inflicted. By 1940, he had begun an affair with Margaret Jennings, a physician and histologist who had joined the Dunn in 1936 and became indispensable to the Australian both as a lab assistant and as the editor of the entire lab’s scientific publications. Which meant that Ethel Florey, who had been assigned to supervise the penicillin team’s clinical trials, was having her papers and reports checked for clarity by her husband’s mistress.*

By comparison, soothing the sensitivities of Ernst Chain was a challenge scarcely worth mentioning. He and E. P. Abraham knew there was no chance of deciphering the structure of penicillin until it could be crystallized—precipitated into something stable enough to be analyzed. Abraham and Chain were still struggling with the structural chemistry when the Lancet article was published. Aware of this, the journal’s editors appended a note reading, “What [penicillin’s] chemical nature is, and whether it can be prepared on a commercial scale, are problems to which the Oxford pathologists are doubtless addressing themselves,” which reads as a bit of an understated scold, as if the Dunn team were keeping at least some of their work secret for now, possibly in order to ensure that they would receive full credit for the discovery.

They were on to something. On September 2, Alexander Fleming paid the Dunn a surprise visit—more a surprise for some of the team than others; Chain evidently thought Fleming was dead—to find out what had been done “with my old penicillin.” The maneuvering for credit was well under way.

Anyone familiar with the adage about success having a thousand fathers might have predicted what would ensue. The Lancet article had ended, courteously enough, with a footnote from the authors thanking the Nuffield Trust, the Medical Research Council, and the Rockefeller Foundation for their support. It did not have the intended effect. In a foreshadowing of the looming disputes about credit and recognition that would attach to the penicillin discovery, Edward Mellanby scolded Florey about showing more gratitude to the Rockefeller Foundation than his own MRC: “I shall be surprised if the Rockefeller Foundation are supporting the work to anything like [the] extent [of the MRC support; he was unaware of the 1939 Rockefeller grant] . . . if you have a good thing in your own country, you might as well give it proper credit and not follow those people who, in cases of research, find it more convenient to give foreigners boosts than their own colleagues. . . .”

Sometime shortly after the article’s publication, Florey received a report from Ernest Gäumann of the Swiss Federal Institute of Technology informing him that he had been approached by the Basel-based concern known as Chemische Industrie Basel, or CIBA, to help purify and manufacture penicillin for the company. More alarming, at least to Florey, was the other news from Switzerland: German researchers were eager to examine any available samples of penicillin.

It was a real dilemma. On the one hand, penicillin promised to be a spectacularly important scientific and medical advance, and one of the canons of twentieth-century science was that such discoveries should be shared as widely as possible. Moreover, there was no legal ground for refusing knowledge about therapies, even when the therapeutic information about penicillin was still the very definition of provisional. On the other hand, Britain was literally fighting for its life. Sharing information about a drug that might accelerate the recovery of wounded German soldiers seemed a lot like giving aid and comfort to the enemy. Florey seems not to have tortured himself very long about competing loyalties. He immediately wrote to Edward Mellanby, saying it was “very undesirable that the Swiss and hence the Germans should get penicillin, and I think it would be well worth while to issue instructions to the National Type Collections not to issue cultures of Penicillium notatum to anyone with [a] possible enemy connection, and to send a letter to [Alexander] Fleming to the same effect.”

Mellanby’s reply: “I sympathize with your position, but I do not see how the Medical Research Council can ask their National Type Collections to restrict their dispatch of special cultures . . . to a neutral country like Switzerland.” He went on: “If the sulphonamide compounds had not proved to be so efficacious, I think you might have had a strong case [but] although I do not doubt that penicillin may prove to be superior to the sulphonamide compounds, I have difficulty in believing that this superiority is so great that national interests dictate the withholding of publication.”

By January 1941, Heatley’s penicillin factory had produced enough penicillin to move from testing the stuff on mice weighing 20 grams or so to 150-pound humans. The purpose was as much to discover whether the compound was dangerous as to test whether it was efficacious. Biologists and pathologists had, after all, discovered dozens of antibacterial compounds that were therapeutically useless because they attacked healthy cells as aggressively as they did pathogens. Florey and the clinical physician he had recruited to administer treatment to human patients, L. J. Witts of the Radcliffe Infirmary, considered who would be the subjects for such a test. Volunteers from Oxford? Someone already at death’s door? The Dunn team decided on the latter. On January 17, Elva Akers, a patient at Radcliffe whose cancer was so far advanced that she was given only a month or so to live, volunteered to receive an injection of a tiny amount of penicillin: 100 milligrams.

Given the dozens of mice who had received weight-comparable doses of the drug with no ill effect, the injection ought to have been safe for a human. It was not. Mrs. Akers almost immediately experienced a high fever accompanied by seizures. The reason, however, wasn’t the penicillin, but the contaminants that the biochemical team had been unable to segregate from the penicillin itself. This was a side effect of Chain and Abraham’s success in separating the penicillin filtrate into different layers; by making one of them relatively pure—up to 80 percent pure—they had also enriched the other layers with a high percentage of impurities, and at least some of them were pyrogens: fever-causing compounds.

Pure penicillin, they quickly learned, was benign. The same separation process that Chain and Abraham were using to decipher its structure could also purify it sufficiently, using a more stringent method of chromatography to eliminate the poisons hitchhiking alongside. Mrs. Akers reacted to a second round of injections with neither fever nor trembling.

Safety, though, wasn’t the same thing as therapeutic value. For the next test, the Dunn team needed someone suffering not from cancer, but infection.

They didn’t need to look far. The previous September, an Oxford policeman named Albert Alexander had been working in his rose garden when he scratched his face on a thorn. The scratch became infected, first just at the site of the injury, but soon the bacteria that had been so abundant in the soil around the Alexander garden—streptococci and staphylococci, at a bare minimum—began to multiply and deposit their own toxins in the victim’s body. By October, his scalp had become obviously infected, and Mr. Alexander was admitted to the Radcliffe Infirmary, where, despite the use of sulfanilamides, the infection spread to his lungs. By February, he had abscesses growing in his torso, on his arms, and in his left eye, which he would soon thereafter lose. When Norman Heatley saw him in early February, he noted in his diary that the constable “was oozing pus everywhere.”

On February 12, 1941, Mr. Alexander was given an intravenous injection of 200 milligrams of penicillin—still, though his physicians wouldn’t know this until much later, less than 5 percent pure—and a follow-up intravenous drip of 100 milligrams every three hours. After a single day, the eight injections had caused a miraculous improvement. Alexander’s fever had vanished, he was no longer discharging pus, his face was no longer swollen, and he was able to eat.

The problem was keeping the penicillin flowing at a rate sufficient to maintain the therapeutic effect; it took Heatley’s machines days to produce the amount of penicillin Alexander needed every hour. The Dunn team had learned during the mice experiments that penicillin was quickly excreted by the kidneys, while still retaining its antibacterial properties, so the doctors set up a procedure for collecting Alexander’s urine after each dose, then carrying it via bicycle from the Radcliffe Infirmary to the Dunn laboratory (a mile and a half each way) in order to extract more of the precious stuff.

Alexander wasn’t the only one who needed it. Another patient, a fifteen-year-old boy named Arthur Jones who had contracted a life-threatening infection after a hip operation, was getting a similar course of treatment. The Dunn physicians hadn’t known how effective penicillin would be, nor did they have a clue how much was required for a therapeutic dose. By the end of February, the supply of penicillin, even the recycled variety, was exhausted. Arthur Jones survived. Albert Alexander, however, died on March 15, 1941.

The stuff worked—when, that is, there was enough of it. However, no British university lab was equipped to produce the kilogram of pure penicillin Florey estimated would be needed for the next round of clinical trials. Neither was any chemical firm in the Commonwealth. France was occupied, Germany, Japan, and Italy enemies. Only one place was left.

After the fall of the Soviet Union, it became something of a cliché to describe the United States as the “world’s lone superpower.” In economic terms, it had already earned the title by 1912. The year that Paul Ehrlich discovered Salvarsan, Germany’s gross domestic product was a bit more than $227 billion, just ahead of the United Kingdom’s $216 billion. That year, the United States economy was larger than both of them combined: $498 billion. By 1940, the GDP of the just-out-of-the-Great-Depression United States was closing in on a trillion dollars a year.

To be sure, American economic dominance wasn’t uniform. Though U.S. steelmakers rolled out 43 million metric tons in 1940, nearly a third of the world’s total (Germany’s 22 million metric tons earned it only a distant second place), American chemical and pharmaceutical firms were minnows next to I. G. Farben’s whale. And while researchers in U.S. universities and industries were well on the way to the dominant position they would assume after the Second World War, Germany’s scientific reputation, particularly in physics and chemistry, was still dramatically higher. Though the Nobel Prize is a notoriously imperfect yardstick of scientific achievement, it isn’t a coincidence that by 1940 Germany had won thirty-three of the science Nobels. Americans had won twelve . . . three of them, in 1934, for the same discovery: pernicious anemia.

In one area, however, the United States was unmatched: Both the sophistication and productivity of America’s agricultural sector was like nothing else in the world. Farms, forests, and ranches still made up nearly 20 percent of the largest national economy in the world.

Which is why, when Warren Weaver of the Rockefeller Foundation visited Oxford on April 14, Florey proposed a visit to the United States, explicitly to find “some American mold or yeast raiser who would undertake a large-scale production of this material for a test, say, 10,000 gallons. . . .” Weaver was convinced almost immediately, and authorized $6,000 in expenses for the trip. Florey needed only one authorization: to leave England. In late April, he wrote to Edward Mellanby asking for help in getting the required wartime exit permits for himself and Norman Heatley, Florey’s choice for an expedition devoted to cultivating penicillin in large quantities. A few days later he received this reply: “I have come to the conclusion that the only way that this most important matter can be pursued is for you and Heatley to go to the United States of America for three months.”

On June 27, 1941, Florey and Heatley left Oxford by car for a trip to a top-secret airfield, where they boarded a Dutch passenger plane and flew to Lisbon, landing seven hours later. There they met representatives of the Rockefeller Foundation, and three days later continued their journey aboard the Pan American Airways’s Dixie Clipper. The flying boat discharged its passengers at the Marine Air Terminal of La Guardia Field on the afternoon of July 2. Within hours, Florey was sitting at the head of a conference table in the Rockefeller Foundation offices in Manhattan, explaining the significance of the penicillin experiments to Alan Gregg, the head of the foundation’s Medical Science Division. To be fair, American awareness of penicillin had preceded Florey and Heatley. The Lancet article had prompted a team at Columbia University College of Physicians and Surgeons to request samples of the compound from Chain and to set up a production line in Manhattan similar to Heatley’s in Oxford. By October 1940, in fact, the Columbia team had produced enough penicillin—still the crude and impure filtrate—to inject two humans with it, even before the Dunn team had done the same with Mrs. Akers. A soil scientist at Rutgers College in New Jersey, Selman Abraham Waksman, started some investigations into P. notatum,* as did researchers at the Mayo Clinic in Rochester, Minnesota.

Nor were America’s small but energetic pharmaceutical firms slow to show interest. Decades before, Parke-Davis had agreed to a cooperative relationship with St. Mary’s Inoculation Department; after reading the Lancet article, Parke-Davis executives asked Almroth Wright whether he or Fleming could help to secure a sample of penicillin from Oxford. (This may explain Fleming’s surprise visit to the Dunn School the preceding September.) Pfizer, a chemical company headquartered in the Williamsburg section of Brooklyn, that made most of its income from producing the preservative and flavoring compound citric acid, also had a small interest in medicines. So did E. R. Squibb, another Brooklyn-based company and a major producer of surgical drugs like ether. Even more interested was the American branch of the German drug company Merck, which had begun cultivation of P. notatum as early as January 1940, and whose president, George Merck, was so well known to Alan Gregg that the Rockefeller Foundation executive proposed a meeting with Howard Florey.

Florey was interested, but he had more immediate business. In July of the preceding year, with the Battle of Britain raging, the Floreys had sent their two children, Paquita and Charles, to New Haven, Connecticut; Howard’s onetime Rhodes Scholar companion John Fulton, now the Sterling Professor of Physiology at Yale, had agreed to take them in for the duration. On July 3, Florey headed to Connecticut, intending to surprise his children* and to meet with some like-minded scientists. Fulton was able to perform a dozen different introductions, but none were more significant than those he brokered between Florey and Ross Harrison, chair of the Executive Committee of the National Research Council, which had been responsible for applying “scientific methods in strengthening the national defense” since 1916. Harrison, in turn, arranged an introduction to Charles Thom, a mycologist in the Department of Agriculture’s Bureau of Plant Industry in Beltsville, Maryland, and the following week, Florey and Heatley headed for Washington to meet him.

Thom already had a long-standing connection to the world of penicillin research. He was the scientist who had originally corrected Fleming’s misidentification of his penicillin-producing fungus—not P. rubrum, as Fleming originally had it, but P. notatum, using a sample sent to him by his old friend Harold Raistrick.* More important, though, than Thom’s previous achievements were his current interests and capabilities. Only a few months earlier, a number of his protégés had been relocated from their own labs in Arlington, Virginia—the War Department had taken the land for building what would become the Pentagon—to the Agriculture Department’s largest midwestern lab. As a direct result of their discussions with Thom, on July 12, Howard Florey and Norman Heatley boarded a train leaving Washington, DC’s Union Station bound for Chicago, where they would make a connection to the Peoria Rocket. Their final destination was the Department of Agriculture’s Northern Regional Research Laboratory, in Peoria, Illinois.

In July 1862, Abraham Lincoln signed the Morrill Act, establishing “at the Seat of Government of the United States, a Department of Agriculture.”* In late 1864, during his last address to Congress Lincoln said, “The Agricultural Department . . . is rapidly commending itself to the great and vital interest it was created to advance. It is precisely the people’s Department, in which they feel more directly concerned than in any other.”

Eight decades later, the United States was a considerably less agrarian society, but the USDA was, if anything, even more important to its prosperity. In addition to its programs promoting American food production and providing credit to American farmers, it funded more than forty experimental stations, research centers where farmers, ranchers, and agronomists worked to improve existing agricultural products and practices and develop new ones; provided ongoing education in agronomy and animal husbandry through its cooperative extension programs; and was the primary enforcer of the provisions of the 1906 Pure Food and Drugs Act, including the penalties for misbranding medicine. Of more immediate relevance to Florey and Heatley, though, were the USDA’s four regional labs, the very top of the pyramid in agricultural research.

The four labs—in addition to the Northern Regional Research Lab in Peoria, the USDA operated an eastern lab in Wyndmoor, Pennsylvania; a southern lab in New Orleans; and a western lab in Albany, California—were responsible over the years for literally thousands of innovations, patented and otherwise, from instant mashed potatoes to wrinkle-free cotton. It seems safe to say, though, that the regional labs’ finest hour can be dated from the arrival of Howard Florey and Norman Heatley on the Peoria Rocket on July 14, 1941.

The Northern Lab looms deservedly large in any history of penicillin—in any history of medicine, really. It was where three of the most urgent objectives in transforming the Dunn discoveries from a laboratory process to an industrial one were achieved: first, the discovery and identification of the most productive strains of Penicillium mold; second, a protocol for accelerating the growth of penicillin-producing mold; and third, improvement of the fermentation technique by which the exudate appeared. In traditional agricultural terminology, they were looking for better seeds, better soil, and better cultivation and harvesting.

Better seeds first. Even before Florey and Heatley arrived in July, the Northern Lab’s chief mycologist, Kenneth Raper, had sent messages to researchers all over the globe (even to the point of enlisting crews in the U.S. Army’s Air Transport Command) requesting that they collect samples of Penicillium mold and send them to Peoria. By early 1941, he had started testing dozens of different strains. The most significant, by far, was found by one of Raper’s lab technicians, a bacteriologist named Mary Hunt, who had been charged with the task of visiting Peoria’s markets in search of moldy fruit and vegetables. In 1943, she hit the jackpot: a cantaloupe infested with a mold so powerful that it would, by the end of the 1940s, be the ancestral source for virtually all of the world’s penicillin.

At roughly the same time he sent Mary Hunt on her tour of Peoria’s fruit stands, Raper had charged another of his subordinates, a microbiologist and mycologist named Andrew Moyer, with finding a better soil: a superior growth medium for the fungus, one that improved the best speed that Heatley’s bedpans had been able to achieve using Czapek-Dox and brewer’s yeast. Serendipitously, he had access to an extremely promising replacement. The Northern Lab had been established explicitly to investigate “industrial uses for the surplus agricultural commodities.” In practice, this meant searching for some commercially valuable way of exploiting the waste products left after the American corn harvest—in 1940, more than 56 million metric tons, much of which was turned into corn flakes, animal feed, sweeteners, and a dozen other commodities. The most significant of these, corn steep liquor, was what was left behind after extracting cornstarch. In weeks, Moyer and Heatley, working together, discovered that corn steep liquor plus sugar increased penicillin production significantly. Actually, more than significantly—a thousandfold. This is not a misprint. Earlier that year, the Dunn team had defined what became known as the “Oxford unit,” the quantity of penicillin that, when dissolved in 1 cc of water, inhibited a standard measure of bacterial growth.* The new growth medium was able to improve production from 2 Oxford units per cc of broth to 2,000.

 Discussing of Work on Penicillin

Credit: Peoria Historical Society

The Northern Lab team, including Andrew Moyer (left side, fifth from left) and Robert Coghill (back table, fourth from left)

This left the harvesting problem: fermentation itself, which was a challenge not just of biology, but geometry. The metabolic process by which sugars are converted to acids, gases, and alcohol had many forms, as had been known even before Pasteur, but the Penicillium mold had, thus far, fermented only on the surface of a growth medium, usually agar (this is why flat Petri dishes were and remain so common in biology experiments). Since surface fermentation meant only two dimensions were available for the growth of the compound, expanding the harvestable quantities of the mold seemed to require very large surfaces—Heatley’s bedpans blown up to the size of basketball courts.

It was Robert Coghill, the chief of the Fermentation Division at the Northern Lab, who first proposed using the same sort of deep fermentation used in brewing beer for growing penicillin.

Deep fermentation wasn’t a completely novel idea. A German-speaking Czech chemist named Konrad Bernhauer had published dozens of papers on the subject from 1920 onward.* Even earlier, as far back as the First World War, Pfizer had been investigating deep (or, as it was then known, “submerged”) fermentation in order to improve yields of what would become their core product: citric acid, the age-old flavoring agent and preservative.

The only way to produce citric acid had been extracting it from fruit, particularly lemons, until the German chemist Carl Wehmer had shown that it was produced by molds as well, specifically Penicillium. However, making citric acid by extracting it from a mold was just as likely to produce oxalic acid, which was both unwanted and dangerous. In 1917, Pfizer’s brilliant research chemist James Currie* discovered that Aspergillus niger was a factory for citric acid—feed it sugar, harvest the acid—and started the program the company christened SUCIAC, Sugar Under Conversion to Citric Acid. Enough citric acid, in fact, that, by 1929, Pfizer was selling more than $4.5 million of it.

Such fermentation was aerobic, with A. niger exposed to air using Heatley-like shallow trays. By 1931, though, Pfizer’s chemists had graduated, if that’s the right word, to production of citric acid in relatively small flasks, about 1 liter in volume, in which a powerful stirrer kept the fluid aerated, a process for which they applied for a patent.

At the time Moyer and Coghill started their researches, the technique had been used only for citric acid. Why not penicillin? As Pasteur had been the first to notice, all forms of fermentation are largely similar. In theory, therefore, the same industrial processes used to manufacture citric acid—or, for that matter, beer—could be enlisted for the new miracle drug. The differences weren’t trivial: Beer and citric acid could be fermented in relatively unclean environments, but the air used by fungal cells to produce penicillin needed to be sterile in order to avoid introducing dangerous impurities; the temperature within the hypothetical vat needed to be kept constant; and the process required some way of keeping the whole mess mixed so that each liter had the same amount of growth medium and mold as every other. But they weren’t insuperable. As early as 1937, the USDA’s By-Products Lab at Ames, Iowa (a predecessor of the Northern Lab), had designed an aluminum rotary fermenter. By the fall of 1941, the Peoria team had a demonstration vat—a drum with a washing machine–like agitator, and an injector through which sterile air could be constantly introduced to the soupy contents. Rotary drums like it would be manufacturing penicillin in industrial quantities for the next five years.

At the same time that the Peoria scientists and engineers were cultivating ever more powerful and pure strains of penicillin, the new drug’s potential was exceeding even the hopes of its most passionate advocates, particularly in Britain. Henry Dawson, at Columbia University College of Physicians and Surgeons, injected two patients with a crude filtrate of penicillin broth, though at such a low concentration that it was both safe and ineffective. Between June and August 1941, five more staph-infected patients in Britain—more volunteers from Oxford’s Radcliffe Infirmary—were treated with it; three of them were children, precisely because the quantity of the drug was so limited, and a child could be expected to respond to a lower dose. On August 16, the Lancet featured another article from the Dunn team that reported “that in all these cases a favourable therapeutic response was obtained . . . ,” though in one—the tragic case of four-and-a-half-year-old John Cox—the penicillin cured the staph infection that had caused septicemia in his sinus orbits, lungs, and liver, but could do nothing about the ruptured spinal aneurysm that killed him two weeks after he started antibiotic treatment.

In September 1941, Florey, having completed his proselytizing in the United States, returned home to England to continue his own investigations, leaving Heatley behind to work at the Northern Lab. He was, in consequence, not on hand when the tiny vial of brown penicillin powder that saved the life of the septicemic Anne Miller in March 1942 had, in some sense, marked not just the birth of the antibiotic age, but also the end of European, and especially British, preeminence in the field. The following month, Florey opened a package from the United States expecting to find his long-awaited kilogram of penicillin. It contained only five grams; less, in fact, than the amount that Merck had rushed to New Haven Hospital to save Mrs. Miller’s life. The Americans were keeping everything they could produce for their own research; despite Florey’s tireless campaigning during the summer and fall of 1941, British research on penicillin still relied on what Britain could produce at home.

While Heatley and Florey were traveling across North America drumming up support for their original request—that elusive kilogram of penicillin needed back in Oxford for clinical testing—the Dunn School had remained the most important antibiotic research center in the world, though still reliably dysfunctional. Chain had learned that his colleagues were leaving for the United States only when he noticed their readied luggage. Not only had he not been invited; he hadn’t even been informed. He was furious.

The rationale—that the purpose of the trip was to promote industrial manufacture of the drug, which was Heatley’s special province, not to discern its structure, which was Chain’s—was strong, but unpersuasive. Though Florey understood, earlier than most, that penicillin would never realize its potential either as a therapy or as a scientific breakthrough until its structure was completely understood, he had misunderstood his émigré colleague. Chain was even more avid for recognition than Florey, and feared that he would be cheated out of “his” Nobel Prize—yes; he was already dreaming of a call from Stockholm—unless he promoted his involvement in the discovery.

He wasn’t just sensitive to slights concerning his scientific importance. He was equally sensitive (probably more understandably) about his status as a Jewish émigré. Though Chain could not yet know the fate of the mother and sister he had left behind—both would die in 1942, likely in the Theresienstadt concentration camp—he was well aware that the world had rarely been so dangerous for Jews. In his own words, “One could not trust any undertaking given by Florey. I gave in in the end . . . to avoid any action which could provoke latent anti-Semitism, which was very widespread. I had to bear in mind the fact that the Jewish community would not be very pleased if controversies of any kind with anti-Semitic overtones came into the open. . . . I have always considered, and still do, that Florey’s behaviour to me in the years 1941 until October 1948, when I left Oxford . . . was unpardonably bad.”

During the first months of 1942, Chain and Abraham finally perfected the chemical technique required to produce a stable penicillin salt. The tiny quantities of the drug still being produced by Heatley’s jury-rigged factories were producing better and better results on patients from the Radcliffe Infirmary, and, at the instigation of Winston Churchill, Britain’s drug companies were finally stepping up to the challenge. By the fall of 1942, Kemball, Bishop was sending 150 gallons of broth to Oxford every ten days, which allowed the Dunn—by far the most productive penicillin “factory” in Britain—to supply sufficient research material to Ethel Florey during 1942 and well into 1943.

Nonetheless, the other Dr. Florey recognized that the center of gravity in penicillin production was shifting irrevocably to the United States, the only place with sufficient penicillin for clinical trials. Florey’s original research program had consisted of three key objectives: finding more potent penicillin strains, increasing manufacturing capacity, and unlocking the compound’s chemical structure. Two of them were rapidly outgrowing England. Peoria was well on its way to identifying the most promising strains of Penicillium, and—as will be seen—American industry was about to be enlisted in manufacturing the precious stuff in quantity. The third goal, however, remained on the Dunn School’s research agenda: discovering penicillin’s elemental components, its structure, and its mechanism of action—how it worked.

As can’t be repeated too often, it’s a great deal easier to discover a compound’s ingredients than to understand how they fit together. Fairly rudimentary tools, for example, can identify the presence of sugar, flour, butter, and eggs in a cake; they will tell next to nothing about how the chains of proteins, carbohydrates, and fats link together. And they tell nothing at all about oven temperature or baking time. So, too, with the chemistry of penicillin. The ultimate goal of all the researchers was a method of producing penicillin in a factory, rather than growing it in fermentation tanks, to ensure both consistent quality and increased quantity. It was the same way that Ehrlich created Prontosil: analysis, then synthesis.

This is not to imply that analysis of penicillin was a trivial matter. Teams on both sides of the Atlantic struggled for more than a year to establish the type and number of elements that composed the penicillin molecule. As early as 1940, the Dunn team had demonstrated that, like so many organic molecules, penicillin always contained carbon, hydrogen, nitrogen, and oxygen. But from the beginning, sulfur had also been appearing in samples of the compound. Some samples, though not all. The inconsistencies could have been due to mistakes in the chemical analysis, or the fact that sulfur was just another impurity that was introduced during the growth or extraction processes.

Or, possibly, sulfur might have been central to its effectiveness. Chain and Abraham, along with other organic chemists from Oxford’s Dyson Perrins Laboratory, were determined to solve the sulfur puzzle. By then, Chain, using a technique known as partition chromatography on silica gel, was able to produce penicillin salts that were between 70 and 90 percent pure, which he could then oxidize—that is, turn into acids. Although what was left behind when penicillin was broken down to simpler compounds—formally “degradation products”—weren’t of any therapeutic value, they did have one extremely important aspect: The degradation products, primarily penicillamine, penillic acid, and the penilloaldehydes, were all crystals.

Crystals could be analyzed.

As far back as the seventeenth century, the polymaths Robert Hooke and Johannes Kepler were independently speculating that crystals like gemstones, common salt, and even snowflakes were structurally similar, each with visibly flat faces meeting at regular angles, all in a repeating series. As scientists realized that all matter is composed of atoms, they also concluded that macroscopic crystals reflected a microscopic structural similarity: The atoms that composed them had to be organized in some regular, and therefore decipherable, structure. Structure would explain, for example, why pure carbon can take the form of both the soft graphite in a wooden pencil and a diamond.

Even as late as the 1920s, crystallography was exiled to the disreputable Department of Mineralogy, which was mostly about taxonomy: classifying all of the earth’s naturally occurring crystals, a task on which the Oxford mineralogist Thomas Vipond Barker spent his career. In 1942, one of his students began working with Chain and Abraham on the analysis of penicillin. Her name was Dorothy Crowfoot Hodgkin, still, as of this writing, Britain’s only female science Nobel laureate.

When Dorothy Mary Crowfoot arrived at Oxford’s Somerville College in 1928—she didn’t marry Thomas Hodgkin until 1937—the science of X-ray crystallography was barely a decade old, though the principle of diffraction had been known for centuries, since the Scottish mathematician James Gregory noticed that rays of sunlight split into a spectrum of colors when passing through a bird’s wing; light-wave spectroscopes had been used to analyze chemical elements ever since. X-rays, on the other hand, which had been discovered by the German physicist Wilhelm Roentgen in 1895, travel in waves that are thousands of times shorter than light waves. This meant that the prisms and other tools used to split light waves were too coarse a filter, about as useful for diffracting X-rays as a fishing net was for catching gnats. In 1912, though, another German physicist, Max von Laue, proposed that the tightly packed and repetitive atoms found in crystals—copper sulfate, for example—could do the job. Three years later, the father and son British physicists William Henry Bragg and Lawrence Bragg described the math that could decode the pattern of myriad dots recorded on a photographic plate after scattering X-rays through a crystal: If the X-ray wavelength and the intensity of the image were known, the molecular structure of the crystal could then be calculated. Table salt, for example, as the Braggs showed, has a formula of one atom of sodium plus one of chloride: NaCl. But the X-ray diffraction pattern shows that it is organized in alternating cubes, an atom of sodium surrounded by six chlorine atoms, next to a single chlorine surrounded by six sodium, repeating one after the other.

 British chemist Dorothy Hodgkin (1910 - 1994) at work, 4th December 1964. Hodgkin was awarded the 1964 Nobel Prize in Chemistry 'for her determinations by X-ray techniques of the structures of important biochemical substances'. (Photo by Harold Clements/Daily Express/Hulton Archive/Getty Images)

Credit: Getty Images/Howard Clements

Dorothy Crowfoot Hodgkin, 1910–1994

Dorothy Crowfoot was barely fifteen, and still living in Khartoum—her father, John Winter Crowfoot, was employed in the Egyptian Education Service of Britain’s Colonial Office—when Dr. A. F. Joseph, a chemist working for Wellcome Laboratories, gave her a book by Sir William Bragg on X-ray diffraction, and so kindled her fascination with crystals. At Oxford, the fascination turned into a passion. Under the tutelage of Robert Robinson, a future chemistry Nobelist and the head of the Dyson Perrins lab, and especially J. D. Bernal, she mastered the technique of using X-ray crystallography to map the internal structure of organic molecules several orders of magnitude more complex than Bragg’s table salt.* The list included cholesterol, testosterone, progesterone, pepsin, and a dozen more, most particularly insulin, whose structure Hodgkin began investigating in 1934, and finally resolved thirty-five years later, in 1969.

By 1940, Hodgkin had begun a relationship even more fruitful and long-lasting than the one with Bernal, accepting her first grant, in the amount of £1,000, from Warren Weaver of the Rockefeller Foundation (though, Weaver, whose eye for talent had already found Howard Florey and Ernst Chain, described her request for a grant “for research on proteins and viruses, with the object of carrying out more general and fundamental investigations . . .” as an “uncertain proposal at the present moment”). It was the beginning of a nearly thirty-year relationship with the Rockefeller Foundation, one of the longest in the foundation’s history.

Also in 1940, her extraordinary talents—and her team at Oxford’s Laboratory of Chemical Crystallography—were recruited to the penicillin project. Even before the publication of the August 1940 article in the Lancet, she had been given a heads-up about the miraculous discoveries going on at the Dunn School, when she encountered Ernst Chain on a walk along Oxford’s South Parks Road, just after the first mice had been successfully treated with penicillin. Chain promised, “Some day we will have crystals for you.” It took more than a year for the promise to be redeemed, but in November 1941 Hodgkin wrote to her husband, “I’ve just come back from visiting Chain and now it’s 10:30 P.M. I’m feeling disgustingly cheerful as a result of my visit. . . . Apparently [penicillin] hasn’t yet been crystallized after all. . . . Chain seemed quite keen to let me have some stuff”—the degradation products—“and I’d simply love to try.”

She quickly found that the degradation products were extremely difficult to work with: tiny crystals that were, in Hodgkin’s words, “immersed in a gummy fluid [and] extremely hygroscopic [i.e., attracting water molecules from the environment], so that they are practically impossible to leave out in the air for more than a few minutes or so.” They did, however, reveal one of penicillin’s secrets: Sulfur was an essential element of the compound, which seemed to be a molecule containing nine atoms of carbon, eleven of hydrogen, four oxygen, two nitrogen . . . and one atom of sulfur.*

But how were the atomic bricks arranged into a structure that was able to stop infectious pathogens cold? X-ray crystallography was a powerful tool for investigating such questions, but not an easy one. Creating the picture formed by the X-rays scattered by penicillamine crystals onto a photographic plate was only the first step, and as likely to be confusing as to be revealing.

The reason is that the intensity of the dots captured on a photographic plate measures the amplitude of the wave of the X-ray as it bounced off the faces of the compound’s crystalline lattice, from which a complete picture of the crystal could, in principle, be derived. More electron density, higher amplitude. But a dot that made a dense, high-intensity impression on the plate could just as easily be the result of two waves that were both arriving at the same frequency—the same number of waves per second—and amplitude. When those kind of signals arrived “in phase” with one another, they reinforced the density. In the same way, a low-density image could be produced by two waves that were out of phase and canceled one another out.

Phases: In the top graphic, two waves are in phase—that is, completely overlapping. In the middle, the waves are out of phase, and, in the bottom, completely out of phase.

Solving the potential confusion demanded a subtle and tricky mathematical technique, one that had originally been developed in the early nineteenth century to describe how waves of heat travel in three dimensions over time. As an example, if a match is held under the center of a coin, the coin’s center will heat up; after the match is extinguished, the heat will dissipate in an ordered way, with the center of the coin getting colder, and the edges warming, until an equilibrium is reached. This phenomenon, however, is an extremely complex business, which is why, in the early nineteenth century, the French mathematician Jean-Baptiste Joseph Fourier helped to develop a technique that turns complicated wave functions like these into a collection of the simple sine or cosine waves that are taught to high school trigonometry students.

Unfortunately, while X-ray amplitudes are relatively easy to chart with a simple molecule—remember the Braggs’ table salt—they rapidly become more complicated as molecules get larger. The electron density photographs start to resemble three-dimensional weather maps, complete with eddies, currents, and whirlpools. Diagramming a molecule the size of penicillin—essential for a full understanding of its structure—and calculating which waves were in or out of phase, would take Hodgkin more than two years.

In the meantime, on both sides of the Atlantic, progress was accelerating in the race to produce penicillin in useful quantities: more animal trials, increasingly sophisticated production methods, and, of course, more precise maps of the mysterious compound. Most of the key work was invisible to the public at large; though, on the heels of the Dunn team’s August 1941 Lancet article, the September 15, 1941, issue of Time magazine reported:

A marvelous mold that saves lives when sulfa drugs fail was described in the British Lancet last month by Professor Howard Walter Florey and colleagues of Oxford. The healing principle, called penicillin, is extracted from the velvety-green Penicillium notatum, a relative of the cheese mold. Although it does not kill germs, the mold stops the growth of streptococci and staphylococci with a power “as great or greater than that of the most powerful antiseptics known.”

The article didn’t really generate much in the way of follow-up. Neither, implausibly, did the cure of Anne Miller in March 1942. From the standpoint of scientists like Heatley, Coghill, Florey, and even Chain, that was as it should be. With barely enough penicillin being produced for the minimum number of human trials, the last thing anyone needed was to raise the hopes of every patient in the English-speaking world. But, as the tide of war against the Axis seemed to slowly shift in favor of the Allied armies, the public, in the United States and Britain particularly, was primed to embrace a lifesaving miracle, the world’s first antibiotic drug.

And, of course, the hero who discovered it.

At the end of August 1942, the Times of London published a brief editorial promoting a greater degree of public investment in penicillin development. Either out of ignorance or delicacy, the editorial declined to lionize (or attack) any scientist or public official by name.

It’s impossible to know what reaction the editorial might have generated on its own. However, on August 31, the Times published the following letter:

Sir,—In the leading article on penicillin in your issue yesterday you refrained from putting the laurel wreath for this discovery around anyone’s brow. I would, with your permission, supplement your article by pointing out that, on the principle of palma qui meruit ferat,* it should be decreed to Professor Alexander Fleming of this research laboratory. For he is the discoverer of penicillin and was the author also of the original suggestion that this substance might prove to have important applications in medicine.

As the careful reader might have guessed from the phrase, “this research laboratory,” the letter was signed:

Almroth E. Wright

Inoculation Department, St. Mary’s Hospital

Fleming’s boss and mentor, who remained one of Britain’s most famous physicians, had spoken. Within hours, Fleet Street emptied out and headed to Paddington to find Fleming and Wright. An interview with Fleming appeared in the Evening Standard of August 31. On September 1, half a dozen other papers reported on Fleming’s discovery. Fleming was named the “Man of the Week” by the News Chronicle. Some stories actually had the lab at St. Mary’s producing the samples used at Oxford. In the Daily Mail, Fleming even gave the very strong impression that St. Mary’s, which had last investigated the properties of penicillin seven years before, was the place where a breakthrough could be expected, saying, “the production of the drug is very complicated and the difficulties are great but they are being overcome.”

The obvious implication—that St. Mary’s was the center of penicillin research—didn’t go uncommented. English organic chemist and Nobel laureate Sir Robert Robinson had already replied to Almroth Wright with his letter to the Times, which appeared on September 1 and which read, in part, that if Fleming should wear the laurel wreath, then “a bouquet, at least, and a handsome one, should be presented to Professor H. W. Florey . . . he, and his team of collaborators, assisted by the Medical Research Council, have shown that penicillin is a practical proposition.”

Florey was glad of Robinson’s support, but angry nonetheless. On December 11, 1942, he wrote to Henry Dale, president of the Royal Society, “I have now quite good evidence, from the director-general of the BBC in fact . . . that Fleming is doing his best to see the whole subject is presented as having been foreseen and worked out by Fleming and that we in this department just did a few final flourishes.”

Public confusion about the discovery of penicillin goes on. Recreations in a BBC documentary from the 1970s include a dramatized Fleming preparing the compound to treat Albert Alexander, and even got the year wrong. W. Howard Hughes’s biography of Fleming, Alexander Fleming and Penicillin, claimed that St. Mary’s technicians had been making penicillin every week since its discovery in 1928. The durability of the resentments expressed by the Dunn School team are, in their way, enlightening: a window onto the motivations of research scientists, where careers can literally rise and fall over a few days difference in announcing even a minor discovery to other professionals. Scientists are human beings, after all, and their reasons for spending thousands of hours in poorly ventilated laboratories for poverty wages are complicated: the thrill of solving difficult puzzles; the joy of exercising their talents cultivated over a lifetime; and, of course, the pride and adulation that are the reward of first discovery.

There are practical consequences to primacy of discovery. As Almroth Wright no doubt realized, the institution that could claim penicillin for its own would be a repository of both public acclaim, and a greater share of the financial resources of philanthropies and government agencies. The ways in which a legitimate claim to discovery profited the discoverers of penicillin—like the discovery of Salvarsan or Prontosil before—remain huge, even without calculating the benefits to society at large.

But those ways pale in comparison to the very tangible profits that would be generated by the discoveries that followed.



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