Studies on purification of methamidophos monoclonal antibodies and comparative immunoactivity of purified antibodies.
- Author:
Su-Qing ZHAO
1
;
Yuan-Ming SUN
;
Chun-Yan ZHANG
;
Xiao-Yu HUANG
;
Hou-Rui ZHANG
;
Zhen-Yu ZHU
Author Information
- Publication Type:Journal Article
- MeSH: Antibodies, Monoclonal; immunology; Chromatography, Affinity; Chromatography, Agarose; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Food Contamination; Fruit; Insecticides; immunology; Organothiophosphorus Compounds; immunology; Pesticide Residues; analysis; immunology; Vegetables
- From: Biomedical and Environmental Sciences 2003;16(2):119-125
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo purify Methamidophos (Met) monoclonal antibodies with two methods and compare immune activity of purified antibodies.
METHODCaprylic acid ammonium sulphate precipition (CAASP) method and Sepharose protein-A (SPA) affinity chromatography method were used to purify Met monoclonal antibodies, UV spectrum scanning was used to determine protein content and recovery of purified antibodies, sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) was used to analyze the purity of purified antibodies, and enzyme-linked immunosorbent assay (ELISA) was used to determine immune activity of purified antibodies.
RESULTSAntibody protein content and recovery rate with CAASP method were 7.62 mg/mL and 8.05% respectively, antibody protein content and recovery rate with SPA method were 6.45 mg/mL and 5.52% respectively. Purity of antibodies purified by SPA method was higher than that by CAASP method. The half-maximal inhibition concentration (IC50) of antibodies purified by SPA to Met was 181.26 microg/mL, and the linear working range and the limit of quantification (LOD) were 2.43-3896.01 microg/mL and 1.03 microg/mL, respectively. The IC50 of antibodies purified by CAASP to Met was 352.82 microg/mL, and the linear working range and LOD were 10.91-11412.29 microg/mL and 3.42 microg/mL, respectively.
CONCLUSIONAntibodies purified by SPA method are better than those by CAASP method, and Met monoclonal antibodies purified by SPA method can be used to prepare gold-labelled testing paper for analyzing Met residue in vegetable and drink water.