Analysis of Dialyzed Flask Technology For Antibody Production

摘要

Efficient and cost effective hybridoma culture is essential to small and large scale monoclonal antibody production for research purposes. This study evaluates multiple aspects of two existing production methods and a new dialyzed cell culture flask method for culturing hybridoma cells. The flask separates the cell cultivation compartment from the cell culture media via a 10kDa cut off limit membrane. This method allows for multiple harvests, longer run times and a super-concentrated supernatant.To determine the advantages of the dialyzed flask technology, an anti-6X histidine epitope tag secreting hybridoma cell line and an anti-AKT3 isoform secreting hybridoma cell line were selected. The current production methods for the anti-6X histidine epitope tag and anti-AKT3 isoform producing clones include an animal method (ascites) and a proprietary suspension culture method. Several criteria were measured including direct labor cost, reagent cost and yield of antibodies. The purified monoclonal antibodies were analyzed by SDS-PAGE 4-20% under denatured conditions and characterized by Western Blot, direct ELISA titration, and immunohistochemistry to assess performance.Monoclonal anti-6X histidine epitope tag and anti-AKT3 isoform from the three methods were affinity purified by protein A. SDS-PAGE analysis showed that all antibody samples had a purity higher than 95%. As indicated by the study, production of monoclonal antibodies in ascites often leads to contamination by endogenous host proteins. Western Blot analysis demonstrated no decrease in performance from the antibodies produced in the dialyzed flask. Immunohistochemistry on prostate cancer slides exhibited similar results from all production methods. An economic analysis comparing the three methods was completed. A significant yield increase and a cost savings were realized from the new production method. This increase in performance was due to the reusable nature of the dialyzed flask and its ability to concentrate the antibodies.