Bristol-myers Squibb Compnay

William V Burnett

Sean M Tonzi

Michael Politino

Thomas J Franceschini

Suo Win Liu

Ross Binder

Joanne L Brown

John J Usher

Shu-Jen Chiang

Guna Romancik

Jonathan D Basch

Bernhard M Schilling

Sivakesava Sakhamuri

Steven S Lee

Linda Matlock

Stephen G Zegarelli

Christoph E Joosten

The present invention describes methods and processes for the production of proteins, particularly glycoproteins, by animal cell or mammalian cell culture, preferably, but not limited to, fed-batch cell cultures. In one aspect, the methods comprise at least two temperature shifts performed during the culturing period, in which the temperature is lower at the end of the culturing period than at the time of initial cell culture. Throughout their duration, the culturing processes of the invention involving two or more downward shifts in temperature sustain a high viability of the cultured cells, and can yield an increased end titer of protein product, and a high quality of protein product, as determined, e.g., by sialic acid content of the produced protein. In another aspect, the methods comprise the delayed addition of polyanionic compound during the culturing period. The delayed addition of polyanionic compound sustains a high viability of the cultured cells, and can extend the growth phase, delay the onset of the death phase, and arrest the death phase.

Mammalian cell culture processes for protein production

The invention provides a glutaryl 7-ACA amidase fromstrain BS-203, which catalyzes the hydrolysis of 7-&#x3b2;-(4-carboxybutanamido)-cephalosporanic acid to 7-aminocephalosporanic acid and glutaric acid. The invention also provides nucleic acid sequences, vectors, and host cells useful in the production of this amidase. The glutaryl 7-ACA amidase can be used for the preparation of 7-aminocephalosporanic acid.

Glutaryl cephalosporin amidase from Pseudomonas diminuta BS-203

The invention provides a glutaryl 7-ACA amidase from strain BS-203, which catalyzes the hydrolysis of 7-&bgr;-(4-carboxybutanamido)-cephalosporanic acid to 7-aminocephalosporanic acid and glutaric acid. The invention also provides nucleic acid sequences, vectors, and host cells useful in the production of this amidase. The glutaryl 7-ACA amidase can be used for the preparation of 7-aminocephalosporanic acid.

The instant invention provides nucleic acid molecules encoding cephalosporin esterase from Rhodosporidium toruloides. In addition, the invention provides isolated cephalosporin esterase, expression vectors, host cells, and methods of producing cephalosporin esterase.

Cephalosporin esterase gene from Rhodosporidium toruloides

Nucleic acids coding for penicillin V amidohydrolase (PVA) from Fusarium oxysporum. Also, expression vectors, host cells and a method for production of PVA.

Penicillin V amidohydrolase gene from Fusarium oxysporum

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Fayetteville, NY, United States of America

William V Burnett