CREATING THE HYBRIDOMA TO PRODUCE MONOCLONAL ANTIBODIES AGAINST RECOMBINANT ANTIGEN DES-GAMMA-CARBOXY-PROTHROMBIN

Abstract

To create a hybrid cell line producing monoclonal antibodies against recombinant Des gamma carboxy prothrombin (DCP) antigen in purebred BALB/c mice. Recombinant DCP antigen was mixed with FIA adjuvant and injected into mice. Lympho B cells were harvested from the spleen and lymph nodes of the mice and fused in DMEM medium supplemented with PEG. On the 9th day of culture, cells reached approximately >80% confluency, and hybridoma cells capable of producing specific antibodies against the recombinant antigen were screened, resulting in the selection of the two best-performing hybridoma cell lines. The antibody titers were evaluated after scaling up the cell numbers, and successfully harvesting the monoclonal antibody-producing hybridoma lines. The concentration of monoclonal antibodies in the harvested supernatant from the two hybridoma lines, DCP1 and DCP2, reached a concentration of 1.73 (mg/ml) for DCP1 and 2.15 (mg/ml) for DCP2. These are important initial successes, aiming to create early diagnosis kits for hepatocellular carcinoma.