Gene amplification and vector engineering to achieve rapid and high-level therapeutic protein production using the Dhfr-based CHO cell selection system |
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Authors: | Jonathan J. Cacciatore Lawrence A. Chasin Edward F. Leonard |
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Affiliation: | 1. Department of Chemical Engineering, Columbia University, New York, NY 10027, United States;2. Department of Biological Sciences, Columbia University, New York, NY 10027, United States |
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Abstract: | Demand is increasing for therapeutic biopharmaceuticals such as monoclonal antibodies. Achieving maximum production of these recombinant proteins under developmental time constraints has been a recent focus of study. The majority of these drugs are currently produced in altered Chinese hamster ovary (CHO) cells due to the high viability and the high densities achieved by these cells in suspension cultures. However, shortening the process of developing and isolating high-producing cell lines remains a challenge. This article focuses on current expression systems used to produce biopharmaceuticals in CHO cells and current methods being investigated to produce biopharmaceuticals more efficiently. The methods discussed include modified gene amplification methods, modifying vectors to improve expression of the therapeutic gene and improving the method of selecting for high-producing cells. Recent developments that use gene targeting as a method for increasing production are discussed. |
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Keywords: | Biopharmaceutical Amplification Methotrexate Expression Enhancers |
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