The therapeutic and diagnostic potential of monoclonal antibodies cannot be realized in the absence of large scale production capabilities. For some human studies, gram quantities of isolated antibodies will be required. The goals of Core C are prepare and purify Ig from hybridomas and transfectomas of interest in sufficient quantities for studies proposed in this program. For cloned/engineered antibodies, the best expression system has proven to be myeloma cells. Techniques for large scale Ig production from hybridomas can be applied to the production of Ig from such transfected myeloma cells (transfectomas). We propose to use a state-of-the-art continuous perfusion cell culture system for large scale Ig production from hybridomas. The same system will be used to optimize Ig production from selected transfectomas of interest. Even though transfectomas yields are predicted to be significantly lower than hybridoma yields, the proposed culture system should enable us to prepare sufficient quantities for the proposed studies. Immunoglobulin will be purified by preparative HPLC in a one or two step procedure involving ion exchange and/or Protein A columns. Purified antibody will be provided to Project 1 for quality assurance testing and distribution to other projects.
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