摘要
目的利用噬菌体随机肽库筛选展示河豚毒素(TTX)模拟抗原表位的噬菌体,并以其替代毒素建立免疫学检测方法。方法以抗TTX单克隆抗体为靶分子,从以融合蛋白形式表达在丝状噬菌体M13外壳蛋白Ⅲ表面的随机七肽库中进行生物亲和淘选,用ELISA法鉴定阳性克隆。结果通过4轮淘选,获得了7株能与抗TTX单克隆抗体特异性结合的噬菌体,采用间接竞争ELISA,筛选到3株能抑制TTX的阳性克隆。其中以4号噬菌体建立的免疫检测方法,线性范围为1～20ng/ml(TTX毒素浓度),R2=0.9947,检测下限为1ng/ml。结论从噬菌体七肽库筛选到了展示TTX模拟抗原表位的噬菌体,淘选出来的噬菌体粒子可作为毒素的替代品用于免疫学检测。
Objective To screen the positive phage displaying the mimic epitope of tetrodotoxin（TTX） by using phage random peptide display library technology and to establish immunoassay for the detection of tetrodotoxin.Methods Monoclonal antibody against TTX was used as a ligand to screen the binding peptide from the Ph.D.-7 peptide library.The library is displayed as a fusion protein with the coat protein Ⅲ of filamentous phage M13.The positive clones were identified by ELISA.Results After four rounds of panning,7 positive phages binding to the anti-TTX monoclone antibody were obtained,and through indirect competitive ELISA,3 positive clones inhibiting TTX were screened.A competitive ELISA was established with phage P4,the linear range of the inhibition is 1 20ng /ml,R2 = 0.9947,the detecting limit is 1ng /ml.Conclusion The phage display technique can be successfully applied to screen the mimic epitope of tetrodotoxin.The acquired phages may be used as the surrogate of the toxin to establish immunoassay.
出处
《卫生研究》
CAS
CSCD
北大核心
2010年第3期299-301,305,共4页
Journal of Hygiene Research
