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Some firms worked out collaborative agreements of their own (e.g., Merck and Squibb in February 1942, joined by Pfizer in September). Merck's pilot plant continued to produce several hundred liters of penicillin amoxicillin 1000 mg uses culture per week using both flasks and tray, and in December, Heatley joined the Merck research staff for several months, where he introduced the Oxford cup plate method of penicillin assay, which soon became a standard method industry-wide.

By March 1942 enough penicillin had been produced under OSRD auspices to treat the first patient (Mrs.

Ann Miller, in New Haven, Connecticut); a further ten cases were treated by June 1942, all with penicillin supplied by Merck & Co., Inc.

Pharmaceutical and chemical companies played an especially important role in solving the problems inherent in scaling up submerged fermentation from a pilot plant to a manufacturing scale.

As the scale of production increased, the scientists at Merck, Pfizer, Squibb and other companies faced new engineering challenges.

Smith captured the complexity and uncertainty facing these companies during the scale-up process: "The mold is as temperamental as an opera singer, the yields are low, the isolation is difficult, the extraction is murder, the purification invites disaster, and the assay is unsatisfactory." Because penicillin needs air to grow, aerating the fermentation mixture in deep tanks presented a problem. When corn steep liquor was used as the culture medium, bubbling sterile air through the mixture caused severe foaming. Squibb solved this problem by introducing glyceryl monoricinolate as an anti-foaming agent.

Submerged fermentation also required the design of new cooling systems for the vats and new mixing technology to stir the penicillin mash efficiently. Lilly was particularly successful in making the mold synthesize new types of penicillin by feeding precursors of different structure. Once the fermentation was complete, recovery was also difficult; as much as two-thirds of the penicillin present could be lost during purification because of its instability and heat sensitivity. Methods of freeze-drying under vacuum eventually gave the best results in purifying the penicillin to a stable, sterile, and usable final form.

The steps of fermentation, recovery and purification and packaging quickly yielded to the cooperative efforts of the chemical scientists and engineers working on pilot production of penicillin.

On March 1, 1944, Pfizer opened the first commercial plant for large-scale production of penicillin by submerged culture in Brooklyn, New York.

Meanwhile, clinical studies in the military and civilian sectors were confirming the therapeutic promise of penicillin.

The drug was shown to be effective in the treatment of a wide variety of infections, including streptococcal, staphylococcal and gonococcal infections.

The United States Army established the value of penicillin in the treatment of surgical and wound infections. Clinical studies also demonstrated its effectiveness against syphilis, and by 1944, it was the primary treatment for this disease in the armed forces of Britain and the United States. The increasingly obvious value of penicillin in the war effort led

the

These firms received top priority on construction materials and other supplies necessary to meet the production goals.

The WPB controlled the disposition of all of the penicillin produced. One of the major goals was to have an adequate supply of the drug on hand for the proposed D-Day invasion of Europe. Feelings of wartime patriotism greatly stimulated work on penicillin in the United Kingdom and the United States.

For example, Albert Elder wrote to manufacturers in 1943: "You are urged to impress upon every worker in your plant that penicillin produced today will be saving the life of someone in a few days or curing the disease of someone now incapacitated.

Create an enthusiasm for the job down to the lowest worker in your plant." As publicity concerning this new "miracle drug" began to reach the public, the demand for penicillin increased.

But supplies at first were limited, and priority was given to military use. Chester Keefer of Boston, Chairman of the National Research Council's

Committee

Keefer had to restrict the use of the drug to cases where other methods of treatment had failed.

Part of his job was also to collect detailed clinical information about the use of the drug so that a fuller understanding of its potential and limitations could be developed. Not surprisingly,

Keefer

In every case the petitioner is told to arrange that a full dossier on the patient's condition be sent by the doctor in charge.

When this is received, the decision is made on a medical, not an emotional basis." Fortunately, penicillin production began to increase dramatically by early 1944.

Production of the drug in the United States jumped from 21 billion units in 1943, to 1,663 billion units in 1944, to more than 6.8 trillion units in 1945, and manufacturing techniques had changed in scale and sophistication from one-liter flasks with less than 1% yield to 10,000-gallon tanks at 80-90% yield.

The American government was eventually able to remove all restrictions on its availability, and as of March 15, 1945, penicillin was distributed through the usual channels and was available to the consumer in his or her corner pharmacy. By 1949, the annual production of penicillin in the United States was 133,229 billion units, and the price had dropped from twenty dollars per 100,000 units in 1943 to less than ten cents.

Most British companies moved over to the deep tank fermentation production of penicillin, pioneered in the United States, after the end of the war to meet civilian needs.

In the United Kingdom, penicillin first went on sale to the general public, as a prescription only drug, on June 1, 1946. In Britain, Chain and Abraham continued to work on the structure of the penicillin molecule, aided by the X-ray crystallographic work of Dorothy Hodgkin, also at Oxford.

The unique feature of the structure, which was finally established in 1945, is the

four-membered

In the same year Alexander Fleming, Howard

Florey

The co-operative efforts of American chemists, chemical engineers, microbiologists, mycologists, government agencies, and chemical and pharmaceutical manufacturers were equal to the challenge posed by Howard Florey and Norman Heatley in 1941. As Florey observed in 1949, "too high a tribute cannot be paid to the enterprise and energy with which the American manufacturing firms tackled the large-scale production of the drug.

Had it not been for their efforts there would certainly not have been sufficient penicillin by D-Day in Normandy in 1944 to treat all severe casualties, both British and American." Antibiotics — Open Access Journal. ( A , a ) represent untreated control cells; ( B , b ), ( C , c ) and ( D , d ) represent cells treated by C-M2, NTPS and CM2 + NTPS, respectively.

Scale bars in ( A – D ) = 1 ?m; scale bars in ( a – d ) = 200 nm. Ratios of SFA/UFA in the cell membrane of Morganella sp. ( A , a ) represent untreated control cells; ( B , b ), ( C , c ) and ( D , d ) represent cells treated by C-M2, NTPS and CM2 + NTPS, respectively.

The live cells were stained green and dead cells stained red. Ratios of SFA/UFA in the cell membrane of Morganella sp. ( A , a ) represent untreated control cells; ( B , b ), ( C , c ) and ( D , d ) represent cells treated by C-M2, NTPS and CM2 + NTPS, respectively.

The live cells were stained green and dead cells stained red. Ratios of SFA/UFA in the cell membrane of Morganella sp.

( A , a ) represent untreated control cells; ( B , b ), ( C , c ) and ( D , d ) represent cells treated by C-M2, NTPS and CM2 + NTPS, respectively.

The live cells were stained green and dead cells stained red.

arvensis EOs, antimicrobials (erythromycin and fluconazole) and of different combinations (antimicrobial/EO and EO/EO) against Streptococcus agalactiae and Candida albicans strains.