生化学と分析生化学

概要

Preparation of a Monoclonal Antibody Against Saikosaponin C and the Establishment of its Enzyme-Linked Immunosorbent Assay

Baoping Qu, Jiayang Sai, Huihua Qu, Shuchen Liu, Jinjun Cheng, Wenchao Shan, Yan Zhao and Qingguo Wang

A monoclonal antibody (MAb) 1E11D8 against saikosaponin c (SSc) was prepared by a cell fusion with splenocytes and hypoxanthine-aminopterin-thymidine-sensitive mouse myeloma SP2/0 cell line. The prepared anti-SSc MAb- 1E11D8 has a novel characteristic which shows high specific for saikosaponin c, saikosaponin d and saikosaponin a, three major oleanane-saponins in Radix Bupleuri. By using anti-SSc MAb-1E11D8, a realiable ELISA was developed for the detection of SSc. The system shows a full measuring range from 156.25 ng·mL-1 to 2500 ng·mL-1 in the case of SSc in indirect competitive ELISA (icELISA). The regression equation was y=-0.283 ln(C) +2.3301 with a correlation coefficient of 0.99. Precision and accuracy of the icELISA method were evaluated by the variations between replicates from well to well (intra-assay) and plate to plate (inter-assay). The values obtained for these parameters were within the normal range (<10%). The recovery rates ranged from 99.82 to 103.59%. The ELISA method was further validated to be of use for surveying SSc in traditional Chinese prescriptions.