Antibodies are used in a whole range of disease fields, such as autoimmunity, cancer and inflammation. The FDA first approved therapeutic antibodies for cancer treatment in 1997. The pharmaceutical industry is continuously developing new and robust discovery platforms for production of novel antibodies as therapeutic agents. In this presentation, we describe the down-stream process of clinical manufacture for the purification of an anti-TAC monoclonal antibody .This antibody was purified from bioreactor culture medium, followed by two step Sepharose columns. The product was collected and loaded onto AB-X antigen affinity column. The product was concentrated by ultrafiltration and loaded onto a S-300 Sephacryl column to obtain the final target antibody. This Humanized anti-TAC monoclonal antibody (Daclizumab) , which is directed against IL-2 receptor, will be studied for the prevention of organ rejection in renal transplant patients. Recently, in 2010, the structural model of the IL-2 receptor epitope binding interface with monoclonal antibody was determined using x-ray crystallography. Protein structure analysis using the RCSB protein database and Swiss PDB viewer software was conducted. This permitted analysis of the CD-25 endodomain of IL-2Rα binding domains [D1 (1-64 AA) and D2 (101-164 AA)] against the immunoglobulin Fab beta barrel binding site. The molecular level analysis reveals structural information of binding interface affinity properties between IL-2Rα against IL-2 antagonist antibodies. As the result, both endodomain binding interface of Daclizumab Fab/IL-2Rα complex (code: 3NFP) and IL-2/IL-2Rα complex (code: 1Z92) display a similar binding epitope .This study also provide Proof of Principle of this immune-suppression drug development.

Structural base analysis of production and purification of humanized anti-TAC monoclonal antibody

2019 Spring National Meeting

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