Genentech, Inc.

F. Hoffmann-la Roche Ag

Woon-Lam Susan Leung

James Robert Swartz

Inn Huam Yuk

Dinesh Baskar

Jenny Hsiung

Avi J Ashkenazi

Robert F Kelley

Zahra Shahrokh

Iphigenia Koumenis

Laura C Simmons

Sarah G Hymowitz

Roger Pai

Mark O'Connell

Inn H Yuk

An apparatus and method to maintain pH within a range conducive for cell growth in a bicarbonate-containing cell culture system without the addition of base. The method relies on the gas transfer characteristics of the bioreactor system to modulate the COtransfer to and from the cell culture such that the pH of the cell culture can be maintained within a desired range.

Method of culturing eukaryotic cells

A process is described for producing a polypeptide heterologous towhereincells comprising nucleic acid encoding the polypeptide are cultured in a culture medium while feeding to the culture medium a transportable organophosphate, such that the nucleic acid is expressed. The polypeptide is then recovered from the cells.

Process for producing Apo2L

Refractile particles containing a heterologous polypeptide as an insoluble aggregate are recovered from bacterial periplasm. The process involves culturing bacterial cells so as to express nucleic acid encoding phage lysozyme and nucleic acid encoding the heterologous polypeptide under separate promoters, disrupting the cells mechanically to release the phage lysozyme so as to release refractile particles from the bacterial cellular matrix, and recovering the released refractile particles from the periplasm. Chloroform is not used in any step and the recovery step minimizes co-recovery of cellular debris with the released refractile particles.

Replicable expression vector for bacterial expression of a mammalian polypeptide

Process for bacterial production of polypeptides

Methods of making Apo-2 ligand and formulations of Apo-2 ligand using divalent metal ions are provided. Such divalent metal ions include zinc and cobalt which improve Apo-2 ligand trimer formation and stability. The crystal structure of Apo-2 ligand is also provided, along with Apo-2 ligand variant polypeptides identified using oligonucleotide-directed mutagenesis.

Methods for making Apo-2 ligand using divalent metal ions

Process for producing recombinant polypeptides via a glycerol phosphate or sugar phosphate feed

Processes are described for recovering heterologous polypeptide from bacterial cells, including the periplasm and cytoplasm. One process involves culturing the bacterial cells, which cells comprise nucleic acid encoding phage lysozyme and nucleic acid encoding a protein that displays DNA-digesting activity, wherein these nucleic acids are linked to a first promoter, and nucleic acid encoding the heterologous polypeptide, which nucleic acid is linked to a second promoter, under certain conditions to produce a broth lysate; and recovering accumulated heterologous polypeptide from the broth lysate. Another process entails culturing bacterial cells that comprise nucleic acid encoding phage lysozyme, gene t, and nucleic acid encoding a protein that displays DNA-digesting activity under the control of a signal sequence for secretion of said DNA-digesting protein, wherein said nucleic acids are linked to one or more promoters, and nucleic acid encoding the heterologous polypeptide and a signal sequence for secretion of the heterologous polypeptide, which nucleic acid encoding the heterologous polypeptide is linked to a another promoter that is inducible, under certain conditions to produce a broth lysate; and recovering accumulated heterologous polypeptide from the broth lysate.

Growing community of inventors

San Mateo, CA, United States of America

Woon-Lam Susan Leung