Methods for enhanced protein production

Abstract

The present invention provides a method of increasing protein production in a cell culture by growing cells that produce the protein (e.g., the growth phase) in a perfusion cell culture to a high cell density (i.e., at least above about 40×106 cells / mL) and then switching to a protein production phase, wherein the cells are cultured in a fed-batch cell culture. The present invention further provides a method for clarifying a protein from a cell culture by adjusting the pH of the cell culture to below neutral pH (i.e., below a pH of 7) and settling the cell culture, such that the cell culture separates to form a supernatant layer and a cell-bed layer, wherein the protein is in the supernatant layer.

Description

CROSS REFERENCE TO RELATED APPLICATION[0001]This application is a divisional of U.S. application Ser. No. 13 / 510,372, filed Jun. 14, 2012, which claims the benefit of International Application No. PCT / U.S. 2010 / 056924, filed on Nov. 17, 2010, which claims the benefit of 61 / 261,886, filed Nov. 17, 2009, the entire contents of all of which are herein incorporated by reference.BACKGROUND OF THE INVENTION[0002]Animal cell culture, particularly mammalian cell culture, is commonly used for the expression of recombinantly produced proteins for therapeutic, prophylactic and diagnostic purposes. Although mammalian cell culture methods are preferred over microbial expression systems (e.g., bacterial or yeast expression systems), because they are better suited to express high molecular weight proteins and proteins having complex steric structures, protein expression levels from mammalian cell culture-based systems are generally considerably lower than those from microbial expression systems. W...

AI Technical Summary

Benefits of technology

This approach enables the production of higher quantities of high-quality protein in a shorter time frame, overcoming the limitations of traditional methods by achieving cell densities and protein yields not previously possible, enhancing the efficiency and effectiveness of protein production in mammalian cell cultures.

Problems solved by technology

Specifically, the maximum cell density achieved using known cell cultures techniques is generally thought to be limited by the presence of particular levels of waste products (e.g., lactate, ammonium, etc.), since it is known that such waste products can be toxic to cell growth when they accumulate to critical concentrations (see, for example, Schumpp, B. et al., J.

This increases the freshness, stability and quality of the protein, in contrast to protein obtained from both perfusion and fed-batch cultures subjected to longer periods of culture, which is thus more susceptible to degradation and generally of poorer quality.

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