期刊
ANALYTICAL CHEMISTRY
卷 88, 期 7, 页码 4092-4098出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.6b00426
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资金
- Ph.D. Programs Foundation of Ministry of Education [20130204110023]
- National Science Foundation [31572556]
- International Scientific and Technological Cooperation and Exchange Program in Shaanxi Province [S2015YFKW0002]
- Key Construction Program of International Cooperation Base in S&T, Shaanxi Province, China [2015SD0018]
Chicken is an ideal model for simplified recombinant antibody library generation. It has been rarely been reported to apply chicken single-chain variable fragments (scFvs) in immunoassays for the detection of antibiotic and chemical contaminants in animal food products. In this study, the scFvs (S-1 and S-5) were isolated from a phage display library derived from a hyperimmunized chicken. The checker board titration revealed that the optimum concentrations of S-1 and S-5 were 0.78 mu g/mL and 0.44 mu g/mL respectively, to obtain OD450 around 1.0 at 5 mu g/mL of Gent-OVA coating concentration. Both S-1 and S-5 exhibited negligible cross reactivity with kanamycin and amikacin. The 50% inhibitory concentration (IC50) of S-1 and S-5 were 12.418 mu g/mL and 14.674 ng/mL respectively. In the indirect competitive ELISA (ic-ELISA), the limits of detection for S-1 and S-5 were 0.147 ng/mL and 0.219 ng/mL respectively. The mean recovery for Gent ranged from 60.91% to 118.09% with no more than 10.35% relative standard deviation (RSD) between the intra-assay and the inter-assay. These results indicate the chicken scFv based ic-ELISA method is suitable for the detection of Gent residue in animal derived edible tissues and milk.
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