Preparation and Identification of Monoclonal Antibody against BPA

摘要

Firstly, BPA structure was modified, then coupling BPA with BSA or OVA to prepare immunogen and coating antigen. Five Balb/C mice were immunized with BPA-BSA. Finally an antibody was prepared and the indirect competitive enzyme-linked immunosorbent assay was founded. Results:(1) The monoclonal antibody belongs to IgG1 subtype and К light chain.(2) The antibody titer is 1: 256000, the most suitable concentration of coating antigen is 2μg/mL, and the optimal dilution of antibody and HRP are 1:16000 and 1:10000 respectively. (3)The linear regression line equation is y = 0.1139x + 0.1046, correlation coefficient is R~2=0.97, the detection limit is 0.911ng/mL and IC_(50) is 2.454×103ng/mL. (4)The monoclonal antibody has high specificity for the cross reactivity with phenol, hydroquinone, and tert-butyl hydroquinone being lower than 0.01%, except ortho-hydroxybenzoic acid 2.1%. (5)The recovery range is 93%～116% and 89%～112% when adding BPA into black samples. (6) When the method was used in real materials to detect BPA residual, the results were proximate to the dates by HPLC.