The Study Of Immunoassay For Bisphenol A Based On The Synthesis Of A Poly-L-lysine-hapten Conjugate As A Coating Antigen

Bisphenol A(BPA) belong to estrogenic substances and effects the development of genital system. It is also associated with prostatic cancer, the disruption of pancreatic function and neurotoxic effects. BPA is an important organic chemical material, which is widely used in the synthesis of epoxy resin and polycarbonate plastic. These kinds of products is widely used as food packaging, tableware, plastic bottles, and so on. BPA can be released and diverted into ecological environment under some condition, it is a potential threaten for the our health. Therefore, it is important to establish an effective method to determine the levels of BPA.In this paper, the derivative of BPA was chosen as the hapten. To prepare immunogens. carbodiimide method was used for coupling BPA hapten with bovine serum albumin(BSA). human serum albumin (HSA), and so on. Poly-L-lysine (PLL) and Ovalbumin (OVA) were was used for the preparation of coating antigens. The immunogens was infected into new Zealand female rabbits, and then the polyclonal antibody against BPA was prepared. The Balb/C mouse were infected with the optimal immunogen, its spleen cells were fused with SP2/0cells to prepare hybridoma. The high quality of cell strains against BPA were screen out after four times of subcloning. The polyclonal antibody and monoclonal antibody against BPA were used to develop the immunoassay for BPA.Enzyme-linked immunosorbent assay(ELISA) and fluorescence immunoassay(FIA) were established under optimized immunoassay conditions using BPA polyclonal antibody. In the ELISA method, the linear range of BPA was from1.3ng/mL to292.2ng/mL, the limit of detection of0.5ng/mL, the recoveries from spiked samples were from86.7%to109.2%. The removal ability of watercress against BPA was studied by means of ELISA. the result indicated the removal effect was obvious. In the FIA method, the linear range of BPA was from2.4ng/mL to1279.5ng/mL, the limit of detection of0.8ng/mL, the recoveries from spiked samples were from89.3%to110.6%. It can be seen that enzyme-linked immunosorbent assayr was more sensitive.Monoclonal antibody against BPA was prepared by inducing ascites in vivo. biotin-avidin amplified enzyme-linked immunosorbent assay was established under optimized immunoassay conditions. The linear range of BPA was from0.9ng/mL to678.7ng/mL. the limit of detection of0.3ng/mL, the recoveries from spiked samples were from84.0%to106.0%. The biotin-avidin amplified system had a lower detection limit and cross-reactivity and was suitable for the detection of BPA residual.