摘要
目的从随机肽库中筛选黄曲霉毒素B1模拟抗原表位，以建立无需黄曲霉毒素B1偶联抗原的免疫学检测方法。方法利用噬菌体展示技术，以抗黄曲霉毒素B1单克隆抗体为靶分子，从随机七肽库中进行生物亲和淘选，ELISA鉴定阳性克隆，通过DNA测序对每个阳性克隆进行定性分析。结果通过4轮结合，扩增循环，获得了能与抗黄曲霉毒素B1单克隆抗体结合的肽，采用间接竞争ELISA，筛选到了4株能抑制黄曲霉毒素B1的阳性克隆子，经DNA测序得到HXXDPXH共有序列，其中X为任意氨基酸。以其中P10噬菌体阳性克隆建立竞争ELISA方法，线性范围为500～2000pg／ml，检测下限为50pg/ml。结论利用噬菌体展示技术筛选到了黄曲霉毒素B1模拟抗原表位，并以其代替黄曲霉毒素B1偶联抗原建立了免疫学检测方法。
Objective To screen mimic epitope from phage-display random peptide libraries as sources of peptides that mimic the binding of aflatoxin B1 to monoclonal antibodies raised against the toxin and establish immunoassay for allatoxin B1 Methods The monoclonal antibody against the aflatoxin B1 was used as ligand to screen the binding peptide from the Ph. D.-7 peptide library with phage display and the specificity of colons were identified by ELISA. The peptide sequences of positive phage clones were determined and analyzed by DNA sequencing. Results After four rounds of panning, the binding peptides were screened from random peptide libraries. Through indirect competitive ELISA, 4 positive clones could inhibited aflatoxin B1 and HXXDPXH was characterized by DNA sequencing, X is random amino acid. Then co competitive ELISA was established for clone h e 10, the linear ran es of the inhibition curves were between 100 pg/mland 2000pg/ml, the detecting limit was 50pg/ml. Conclusion The mimic epitope of aflatoxin B1 antigen was obtained successfully by the phage display technigue and it is effective substitutes for aflatoxin B1-protein conjugate in the immunoassay.
出处
《卫生研究》
CAS
CSCD
北大核心
2007年第1期59-62,共4页
Journal of Hygiene Research
基金
"十五"国家重大科技专项课题(No.2001BA804A20)
