Preparation of monoclonal antibody and establishment of indirect competitive chemiluminescence enzyme immunoassay for sodium pentachlorophenolate

Abstract

In this study, three haptens of sodium pentachlorophenolate (PCP-Na) were designed and synthesized, and the monoclonal antibody against PCP-Na with high specificity and affinity was successfully prepared. Based on this monoclonal antibody, a highly sensitive indirect competitive chemiluminescence enzyme immunoassay (ic-CLEIA) was developed for the determination of PCP-Na and its metabolite pentachlorophenol (PCP). The ic-CLEIA method based on optimal conditions showed that the half maximal inhibitory concentration (IC50), linear range (IC20-IC80) and the detection limit (LOD, IC10) for PCP-Na were 0.083 ng/mL, 0.0076–1.5111 ng/mL, and 0.0032 ng/mL, respectively, and 0.29 ng/mL, 0.0983–0.7008 ng/mL, and 0.0192 ng/mL for PCP, respectively. The results of cross reaction showed that the method had no obvious cross reaction with other six chlorophenols compounds (phenol, 2-chlorophenol, 4-chlorophenol, 2, 4-dichlorophenol, 2, 4, 5-trichlorophenol, 2, 4, 6-trichlorophenol) except the metabolite PCP. The average recoveries in chicken, pork and fish samples were 81.3–105.8 %, with coefficient of variation (CV) lower than 7.6 %. In this study, a new highly rapid, sensitive and accurate ic-CLEIA method was established, which can be used for the sensitive detection of PCP-Na and its metabolite PCP in animal tissues.