Momenta Pharmaceuticals, Inc.

Biogen Idec Ma Inc.

Holly Prentice

Brett Belongia

Rasheed Tijani

Brian Edward Collins

William Sisk

Barbara Ehrenfels

William P Sisk

Polypeptides having target levels of C-terminal variants are described.

Cell culture process

Polypeptide preparations having target levels of glycans, and methods of producing such polypeptide preparations using putrescine, are described.

Methods of cell culture

Polypeptide preparations having target levels of glycans, and methods of producing such polypeptide preparations using DMSO, are described.

Polypeptide preparations having target levels of glycans, and methods of producing such polypeptide preparations using ammonium and/or lysine, are described.

A method for producing recombinant polypeptides, including antibodies, having targeted levels of carboxyl terminal (C-terminal) lysine or arginine residues is presented. The method involves culturing cells expressing said polypeptides in cell culture medium having sufficient levels of lysine, arginine, and/or agents that change intracellular pH. Culturing the cells under such conditions does not affect cell growth, cell viability, or titer.

The invention relates to methods for producing mammalian cells having enhanced characteristics such as faster growth rates and the ability to grow to higher cell densities. The invention also relates to methods for producing cells having the ability to grow in media lacking components such as insulin and insulin substitutes. The invention also relates to isolated mammalian cells having improved growth characteristics and/or the ability to grow in media lacking components such as insulin and insulin substitutes.

Methods for producing mammalian cells

The invention provides a method of producing multiple polypeptides, such as antibodies or antibody fragments, in a eukaryotic cell using a single expression vector. The expression vector is engineered to comprise two or more expression cassettes under the control of a single promoter wherein the expression cassettes have splice sites which allow for their alternative splicing and expression as two or more independent gene products at a desired ratio. Use of the vector for the efficient expression of recombinant antibodies in eukaryotic host cells is disclosed as well as the use of such antibodies in diagnostic and therapeutic applications.

Methods and constructs for expressing polypeptide multimers in eukaryotic cells using alternative splicing

The disclosure provides an expression cassette and a vector comprising the cassette for expression of a polynucleotide. The expression cassette includes a promoter/enhancer, an intervening region, and a polyadenylation signal domain. Expression systems and methods of using the expression cassette and vector are also provided.

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Carlisle, MA, United States of America

Holly Prentice