Hapten design and indirect competitive immunoassay for parathion determination: correlation with molecular modeling and principal component analysis.
A novel procedure for parathion hapten design is described. The optimal antigen for parathion was selected after molecular modeling studies of six types of potentially immunizing haptens with the aim to identify the best mimicking target analyte. Heterologous competitive indirect enzyme-linked immunosorbent assay (ELISA) was developed after screening a battery of competitors as coating antigens. The relationship between the heterology degree of the competitor and the resulting immunoassay detectability was investigated according to the electronic similarities of the competitor haptens and the target analyte. Molecular modeling and principal component analysis were performed to understand the electronic distribution and steric parameters of the haptens at their minimum energetic levels. The results suggested that the competitors should have a high heterology to produce assays with good detectability values. An indirect competitive ELISA was finally selected for further investigation. The immunoassay had an IC50 value of 4.79 ng mL(-1) and a limit of detection of 0.31 ng mL(-1). There was little or no cross-reactivity to similar compounds tested except for the insecticide parathion-methyl, which showed a cross-reactivity of 7.8%.
Liu
Yi Hua
YH
Institute of Pesticide and Environmental Toxicology, Zhejiang University, Hangzhou 310029, China.
Jin
Mao Jun
MJ
Gui
Wen Jun
WJ
Cheng
Jing Li
JL
Guo
Yi Rong
YR
Zhu
Guo Nian
GN