الفهرس 
INTRODUCTIONOnly 14 pages are availabe for public view
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Abstract
During the last few decades, monoclonal antibodies (MAbs) produced from hybridoma cell lines have been used in many areas of biomedical research and immunotherapeutic regimens that should be of absolute purity. The aim of the present study is to describe the method for production, purification and different methods for monitoring the purity. For this purpose a mouse hybrid cell line (CRL 8001, ATCC, USA) secreting mouse IgG2a specific to CD3 antigenic determinant expressed by human peripheral T cells was used. The purification, from the clarified culture supernatant, was carried out using industrial media on a process development scale. The process involves two liquid chromatographic steps: affinity chromatography on protein A Sepharose 4 Fast Flow and anion exchange chromatography on Q Sepharose Fast Flow. High performance liquid chromatography (HPLC) analysis and sodium¬dodecyl sulphate¬polyacrylamide gel electrophoresis (SDS¬PAGE) were performed to monitor the purity of the antibody. Absolute purity was obtained in the mouse IgG2a specific fractions. This obvious purity was confirmed using immunoelectrophoresis. Enzyme linked immunosorbant assay (ELISA) was used to quantify the antibody in the different purification stages. The in vivo potency was evaluated depending on a renal transplantation model of outbreed Sprague Dawely rats. Serum creatinine, as a biochemical measure indicating grafted kidney function, was measured. The histological parameters of tissue rejection were studied in counterstained paraffin sections. The results indicate that the purification process was reasonably fast and easy and the recovery was found to be 85% 7.26. Accordingly, the purification procedure described appears to compare favorably with the minimum requirement of purity for human use and that of recovery for scaling up. The MAb was found to be effective in prevention of graft rejection and reversal of acute rejection episode as indicated by normal values of their serum creatinine levels. Rats treated with the MAb, starting from the day of transplantation lived longer than non¬treated rats. Also, rats treated after three days of transplantation were capable to survive longer than non¬treated. Accordingly the MAb can be used in clinical trials.