Monoclonal antibody produced by heterologous indirect enzyme-linked immunosorbent assay and its application for parathion residue determination in agricultural and environmental samples.
A heterologous indirect enzyme-linked immunosorbent assay (ELISA) was developed, which was based on monoclonal antibody (Mab) to determine parathion residue in agricultural and environmental samples. Eight Mabs were produced by introducing the heterologous indirect ELISA into the screening procedure. It was shown that these Mabs had more broad-reactivity with twenty competitors than that of 5H7 (Mab produced by homologous screening). So it became much easier using these new Mabs to develop heterologous immunoassays. In addition, all the developed heterologous ELISAs could be used to determine parathion residue in semiquantitative or quantitative level, and their detection limitation (LOD) was around 2 ng/mL. Compared with the previous heterologous ELISA (hapten 11/5H7) with IC(50) of 13.3 ng/mL, a better heterologous ELISA (hapten 11/1E1) was obtained with IC(50) of 3.8 ng/mL, which improved the sensitivity 3.5 times. Finally, the latter was applied to parathion residue determination in spiked agricultural and environmental samples without extraction or cleanup. The average recoveries of parathion added to water, soil, cucumber, Chinese cabbage and carrot were between 80.4 % and 111.8 %. The LOD for water and soil samples was 5 ng/mL, and the LOD for cucumber, rice and corn samples was 10 ng/mL.
Liu
Yi H
YH
Institute of Pesticide and Environmental Toxicology, Zhejiang University, Hangzhou, China.
Wang
Chun M
CM
Guo
Yi R
YR
Liang
Xiao
X
Gui
Wenjun
W
Zhu
Guo N
GN
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000096:biosynthesis / Q000276:immunology; D002459:Cell Fusion; D002478:Cells, Cultured; D018556:Crops, Agricultural / Q000737:chemistry*; D004784:Environmental Monitoring; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006241:Haptens / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D006825:Hybridomas; D020128:Inhibitory Concentration 50; D057230:Limit of Detection; D008214:Lymphocytes / Q000166:cytology / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009101:Multiple Myeloma / Q000276:immunology; D010278:Parathion / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D010573:Pesticide Residues / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D013154:Spleen / Q000166:cytology / Q000276:immunology
D000911:Antibodies, Monoclonal; D006241:Haptens; D010573:Pesticide Residues; D010278:Parathion