tailieunhanh - Production of monoclonal antibody to quantify serum CA125 concentration in breast and ovarian cancer patients

In this study, we developed a mAb against CA125 to be used as a capture antibody in ELISA technique for detecting CA125 concentration in sera of OC and BC patients. The results of CA125 level in this study were compared with that performed using a commercial CA125 ELISA kit. | ACADEMIA JOURNAL OF BIOLOGY 2020 42 4 109 116 DOI 2615-9023 PRODUCTION OF MONOCLONAL ANTIBODY TO QUANTIFY SERUM CA125 CONCENTRATION IN BREAST AND OVARIAN CANCER PATIENTS Nguyen Thi Xuan1 2 Nguyen Trong Ha3 1 Institute of Genome Research VAST Vietnam 2 Graduate University of Science and Technology VAST Vietnam 3 103 Hospital Vietnam Military Medical University Ha Noi Vietnam Received 27 May 2020 accepted 20 September 2020 ABSTRACT Ovarian carcinoma OC and breast cancer BC are mainly caused by alterations in genes such as BRCA1 and BRCA2 which are involved in differentiation and survival of cancer cells. The protein CA125 MUC16 is released by cancer cells in most OC and a few BC patients. The cut- off point of CA125 level for tumor growth and metastasis is 35 U mL. Production of CA125 monoclonal antidody mAb to determine expression level of this antigen by ELISA has been well known. In this study we aimed to generate CA125-specific mAb for developing a new in- house ELISA kit. To this end BALB c mice were immunized with CA125 protein and splenocytes of immunized mice were fused with myeloma Sp2 0 cells. Efficiency of mAbs secreted from the hybridoma clones was examined by ELISA and flow cytometry analysis. As a result among 3 stable hybridoma cell lines identified A1 clone attained about 90 positive for anti-CA125 mAb whereas H1 and H3 clones were about 40 and 50 positive for anti-CA125 mAb respectively. By flow cytometry analysis anti-CA125 mAb from A1 clone was more specific to CA125 antigen present in OVCAR -3 cells than those from H1 or H3 clone. In addition the isotype of the obtained mAb was specific IgG1 and Kappa light chain. In conclusion the mouse anti-human CA125 mAb generated in our lab was specifically binding to CA125 antigen and used as the capture antibody in sandwich ELISA system for early diagnosis as well as monitoring therapeutic response in OC patients. Keywords CA-125 breast cancer ELISA ovarian cancer monoclonal .