Tag: <span>igg</span>

Hybridoma cell lines are highly unpredictable and often, an unreliable source of important antibodies of national security interest. There is an urgent need to convert antibody production from these cell lines into robust, recombinant platforms that can reliably produce large quantities of antibody on demand, and abandon methods based on murine ascites production. This work describes the use of lentiviral vectors, Chinese hamster ovary (CHO)-K1 cells, and high-density perfusion cultures for antibody production. Cell line development was rapid, high insertion copy numbers were achievable, and the heavy/light chain ratios could be rapidly optimized. The mammalian cells provided an appropriate environment for IgG folding and obviated the difficult purification steps such as removal of endotoxins, refolding, or dealing with abnormal post-synthetic modifications common to other production systems. We found that even in the absence of an optimized cell line, in high-density cultures, routine productivities in the 1–5 g/L range were achieved. As expected, productivity was independent of the performance of the original hybridomas. We conclude that the lentiviral vector system can achieve high copy numbers of immunoglobulin genes with optimized heavy and light chain ratios to appropriately assemble and secrete the immunoglobulins, achieving high productivity. This observation suggests that substantial advances can be made by selecting and optimizing the cell line used for immunoglobulin production. The lentiviral vector-based method of antibody production offers substantial improvements over traditional murine ascites-based antibody production in terms of reliability, productivity, and cumulative cost—if the antibody need exceeds 1–3 grams within the shelf life of the product…