Production of recombinant anti-human CD19 monoclonal antibody express by genetically CHO-DG44 cells

Abstract

Due to the impact of living conditions, the incidence of cancer is increasing day by day. In recent years, every year Vietnam discovers a lot of new cancer patients such as lung cancer, breast cancer and leukemia, … CD 19 is thought to be a promising target for immunotherapies focused on B-cell malignancies. In the flow cytometry technique to identify B cells, in addition to the FACS device, the CD19 marker specific monoclonal antibody is an indispensable material. All anti-human CD19 antibodies used in diagnosis and research are imported biological products with high cost and need more time to deliver. With the high demand for current and future use of antihuman CD19 monoclonal antibodies in disease diagnosis and research. We want to create a low-cost monoclonal antibody system for future studies based on commercial antibody imitation and using pFUSE and pOptiVEC vectors for expression. In this study, based on the published sequence on reliable data, we selected and mimic to generate recombinant plasmid encoded anti-human CD19 monoclonal antibody, transformation into E Coli, screening of E. coli lines carrying target gene and expressing it from CHO-DG44 cells. The pFUSE vector containing the antibiotic resistance gene Zeocin or Blasticidine is used with the expectation that the recombinant plasmid will have a higher expression level and more specificity. The pOptiVEC vector carrying the gene encoding the DHFR protein, the recombinant plasmid will be able to simultaneously express DHFR – protein required to growth CHO cells on nucleoside deficient medium MEM (-) and monoclonal antibodies. Since then, the combination of two recombinant plasmids with the hope of higher expression levels in both intracellular and extracellular. And we may be able to identify the right ratio of light chain and heavy chain plasmids to co-transform into CHO-DG44 cells in order to generate extracellular anti-human CD19 monoclonal antibody