Studies on purification of methamidophos monoclonal antibodies and comparative immunoactivity of purified antibodies / 生物医学与环境科学(英文)
Biomedical and Environmental Sciences
; (12): 119-125, 2003.
Article
em En
| WPRIM
| ID: wpr-264286
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WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To purify Methamidophos (Met) monoclonal antibodies with two methods and compare immune activity of purified antibodies.</p><p><b>METHOD</b>Caprylic 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.</p><p><b>RESULTS</b>Antibody 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.</p><p><b>CONCLUSION</b>Antibodies 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.</p>
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Base de dados:
WPRIM
Assunto principal:
Compostos Organotiofosforados
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Verduras
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Ensaio de Imunoadsorção Enzimática
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Resíduos de Praguicidas
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Contaminação de Alimentos
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Cromatografia de Afinidade
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Cromatografia em Agarose
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Eletroforese em Gel de Poliacrilamida
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Alergia e Imunologia
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Frutas
Idioma:
En
Revista:
Biomedical and Environmental Sciences
Ano de publicação:
2003
Tipo de documento:
Article