Genentech, Inc.

Dana Christian Andersen

Bradley Richard Snedecor

Inn Huam Yuk

Martin Gawlitzek

Tiffany M Bridges

Cynthia A Hoy

Laura C Simmons

John C Joly

A method of reducing glycation of an amino acid in a protein produced by a host cell in cell culture medium is disclosed.

Methods of protein production

The present invention provides methods and compositions for expression and production of recombinant antibodies in a host cell system, such as prokaryotic and eukaryotic expression systems. Particularly contemplated are recombinant systems for temporally separated expression of light chain and heavy chain of antibodies. The antibody products including antibody fragments can be used in various aspects of biological research, diagnosis and medical treatment.

System for antibody expression and assembly

The invention provides a method for increasing product yield of a polypeptide of interest produced by recombinant host cells, where expression of the polypeptide by the recombinant host cells is regulated by an inducible system. More specifically, the method involves culturing the recombinant host cells under conditions of high metabolic and growth rate and then reducing the metabolic rate of the recombinant host cells at the time of induction of polypeptide expression. In particular, the invention provides a method of increasing product yield of an antibody, growth factor, or protease produced by a recombinant host cell regulated by an inducible system.

Metabolic rate shifts in fermentations expressing recombinant proteins

A glycoprotein is produced by a process comprising culturing mammalian host cells expressing nucleic acid encoding a glycoprotein in the presence of (a) a factor that modifies growth state in a cell culture, (b) a divalent metal cation that can adopt and prefers an octahedral coordination geometry, and/or (c) a plasma component. In this process, the occupancy of an N-linked glycosylation site occupied only in a fraction of a glycoprotein is enhanced. Such culturing is preferably carried out at a temperature of between about 30&deg; C. and 35&deg; C. and/or in the presence of up to about 2 mM of a butyrate salt and/or in the presence of a cell-cycle inhibitor.

Cell culture process

A glycoprotein is produced by a process comprising culturing mammalian host cells expressing nucleic acid encoding said glycoprotein in the presence of (a) a factor that modifies growth state in a cell culture, (b) a divalent metal cation that can adopt and prefers an octahedral coordination geometry, and/or (c) a plasma component. In this process, the occupancy of an N-linked glycosylation site occupied only in a fraction of a glycoprotein is enhanced. Such culturing is preferably carried out at a temperature of between about 30&deg; C. and 35&deg; C. and/or in the presence of up to about 2 mM of a butyrate salt and/or in the presence of a cell-cycle inhibitor.

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Menlo Park, CA, United States of America

Dana Christian Andersen