An immunoassay for bisphenol A based on direct hapten conjugation to the polystyrene surface of microtiter plates

摘要

Based on direct hapten coated format a competitive indirect enzyme-linked immunosorbent assay (ciELISA) for bisphenol A (BPA) was developed. Polystyrene surface was modified by 3-Aminopropyltriethoxysilane (APTES) to produce amino groups after H2SO4/HNO3-pre-treatment. 4,4-bis (4-hydroxyphenyl) valeric acid (BVA) which is analogue of BPA, was successfully immobilized on the surface of microtiter plates by N,N'-dicyclohexylcarbodiimide (DCC) method. The essential steps of the assay were optimized, especially blocking procedure which is key step to prevent unspecific binding of antibody. The results indicated that compared with hapten-protein coated format (IC50 = 176.67ng ml(-1), LOD = 15.90 ng ml(-1)), the direct hapten coated format (IC50 = 23.50 ng ml(-1), LOD = 0.27 ng ml(-1)) could improve assay sensitivity and the detection ranges were 2.30 ng similar to 157.60 ng ml(-1) with good signal reproducibility (P value>0.05) after careful optimization of assay conditions. Tap water samples and seawater samples were spiked with a known amount of BPA and measured by ciELISA. The average recoveries were between 70 and 142%. As far as we are aware this is the most sensitive ELISA for BPA yet reported.