Development of Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assay for Gossypol Analysis in Cottonseed Meals
A monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed for the analysis of gossypol in cottonseed meals. First, the checkerboard method was used to determine the optimum amount of coating antigen gossypol-BSA (bovine serum albumin) and primary ant...
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Published in | Journal of agricultural and food chemistry Vol. 52; no. 26; pp. 7793 - 7797 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
Washington, DC
American Chemical Society
29.12.2004
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Subjects | |
Online Access | Get full text |
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Summary: | A monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed for the analysis of gossypol in cottonseed meals. First, the checkerboard method was used to determine the optimum amount of coating antigen gossypol-BSA (bovine serum albumin) and primary anti-gossypol monoclonal antibody (Mab) needed in the ic-ELISA. Second, the effects of several physical (incubation time and temperature) and chemical (solvent types and concentrations) conditions on the performance of Mab on ic-ELISA were investigated to get a rapid robust assay with high sensitivity. Under the established optimized condition, the concentration of gossypol giving 50% reduction of the maximum ELISA signal (I50) in the competitive standard curve was 0.20 μg/mL, whereas the detection limit for gossypol was 0.024 μg/mL. This ic-ELISA method for the analysis of gossypol extracted by methanol from a variety of cottonseed meals was further compared with the official method of the American Oil Chemists' Society (AOCS). The amounts of gossypol determined by the ic-ELISA had a good correlation with those obtained by the AOCS method (R 2 = 0.90). Keywords: Gossypol; cottonseed; monoclonal antibody; immunoassay; ELISA |
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Bibliography: | http://hdl.handle.net/10113/8309 ark:/67375/TPS-1B6XC95P-Q istex:3210C1C28BE725468FE6A2AF417F82AF03B3F339 ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2 |
ISSN: | 0021-8561 1520-5118 |
DOI: | 10.1021/jf049071y |