Development of Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assay for Gossypol Analysis in Cottonseed Meals

• Published in: Journal of agricultural and food chemistry Vol. 52; no. 26; pp. 7793 - 7797
• Main Authors: Wang, Xi, Chen, Feng, Wan, Peter J, Huang, Guohui
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 29.12.2004
• Subjects: Antibodies, Monoclonal; Biological and medical sciences; cottonseed meal; Cottonseed Oil - chemistry; enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - methods; feed contamination; feeds; Fish and seafood industries; Food industries; Fundamental and applied biological sciences. Psychology; gossypol; Gossypol - analysis; indirect competitive enzyme-linked immunosorbent assay; monoclonal antibodies; Solvents; Bass; Edible fish; Meal; cottonseed; Monoclonal antibody; Enzyme immunoassay; Gossypol; immunoassay; Assay; Development; Detection; ELISA assay; ELISA
• DOI: 10.1021/jf049071y
• ISSN: 0021-8561

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