关键词: 生物化学工程, 单克隆抗体, 人参皂苷Rb1, 酶联免疫吸附测定(ELISA)

Abstract: Crosslinking antigens GRb1BSA (ratio 10∶1) and GRb1OVA (ratio 8∶1) were produced by the reactions of ginsenoside Rb1 with serum albumin (BSA) and ovalbumin (OVA) respectively. Three BALB/C mice were immunized five times with GRb1BSA as the immunogen. Using polyethylene glycol (PEG) method, the immunized splenocytes were isolated and fused with a hypoxanthineaminopterinthymidinesensitive mouse myeloma cell line, SP2/0. Hybridomas that produce monoclonal antibody (McAb) reactive to GRb1 were cloned by limiting dilution method. Hybridoma cell line that secretes McAb was obtained. Indirect enzymelinked immunosorbent assay (ELISA) was applied to determine the titer of antisera and competitive ELISA was used to detect McAb. As results, the titer of the antisera is over 1∶16 000 by ELISA, two cell lines (1F7, 1G3) that secrete McAb are obtained, McAb detection shows linear relation in the range from 0.01 g/mL to 1 μg/mL. The full range of GRb1 detection is from 50 ng/mL to 350 ng/mL by competitive ELISA. The two McAbs are contraposed the same determining cluster and can be used to establish ELISA for GRb1 content detection and GRb1 isolation.

Key words: biochemistry engineering, monoclonal antibody, ginsenos ide Rb1, enzyme-linked immunosorbent assay(ELISA)