Preparation of Artificial Antigen and Egg Yolk-derived Immunoglobulin(IgY) of Citrinin for Enzyme-Linked Immunosorbent Assay  

作　　者：ZHAO-HUI DUAN ZHUANG-SEN LIN HE-RUI YAO YAN-HONG GAO KUN ZHANG Su-QING ZHAO ZHEN-YU ZHU 

机构地区：[1]Laboratory of Clinical Immunology, the Second Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510120, Guangdong, China [2]Department of Pharmacy Engineering, Faculty of Chemical Engineering and Light Industry, Guangdong University of Technology, Guangzhou 510006, Guangdong, China [3]Guangdong Center for Disease Control and Prevention, Guangzhou 510300, Guangdong, China [4]Department of Biochemistry, Zhongshan Medical College, Sun Yat-Sen University, Guangzhou 510080, Guangdong, China

出　　处：《Biomedical and Environmental Sciences》2009年第3期237-243,共7页生物医学与环境科学（英文版）

基　　金：supported by the National Natural Science Foundation of China (No. NSFC. 20872020);the Natural Science Foundation of Guangdong Province (No. 06012298 and No. 8251009001000005)

摘　　要：Objective To prepare artificial antigens and anti-citrinin egg yolk-derived immunoglobulin （IgY） to build an enzyme-linked immunosorbent assay （ELISA） for citrinin （CTN）. Methods CTN was conjugated with bovine serum albumin （BSA）, ovalbumin （OVA） with formaldehyde condensation method to prepare artificial antigens and identified by ultraviolet （UV） spectrometry and Infrared （IR） spectrometry. Artificial antigens for CTN and anti-CTN IgY were purified with polyethylene glycol two-step precipitation method and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE）. ELISA with IgY was established. Cross-reactivity of IgY with various structural similarities to CTN and possible co-occurrence with CTN in agricultural commodities were studied. Results UV and IR absorption spectra suggested that CTN was correlated with the carrier protein of BSA or OVA. SDS-PAGE patterns showed that the anti-CTN IgY was almost pure with a molecular weight of approximate 100 KD. The indirect competitive ELISA showed that the detection limit of CTN was 10 ng·mL^-1, with a good linearity ranging 20-640 ng·mL^-1. Conclusion Artificial antigens of CTN can be successfully synthesized. The established ELISA can be used to determine CTN- contaminated samples.Objective To prepare artificial antigens and anti-citrinin egg yolk-derived immunoglobulin （IgY） to build an enzyme-linked immunosorbent assay （ELISA） for citrinin （CTN）. Methods CTN was conjugated with bovine serum albumin （BSA）, ovalbumin （OVA） with formaldehyde condensation method to prepare artificial antigens and identified by ultraviolet （UV） spectrometry and Infrared （IR） spectrometry. Artificial antigens for CTN and anti-CTN IgY were purified with polyethylene glycol two-step precipitation method and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE）. ELISA with IgY was established. Cross-reactivity of IgY with various structural similarities to CTN and possible co-occurrence with CTN in agricultural commodities were studied. Results UV and IR absorption spectra suggested that CTN was correlated with the carrier protein of BSA or OVA. SDS-PAGE patterns showed that the anti-CTN IgY was almost pure with a molecular weight of approximate 100 KD. The indirect competitive ELISA showed that the detection limit of CTN was 10 ng·mL^-1, with a good linearity ranging 20-640 ng·mL^-1. Conclusion Artificial antigens of CTN can be successfully synthesized. The established ELISA can be used to determine CTN- contaminated samples.

关 键 词：CITRININ Artificial antigen lgY ELISA 

分 类 号：R446.62[医药卫生—诊断学] R392-33[医药卫生—临床医学]