利用噬菌体肽库淘选玉米赤霉烯酮的模拟表位

Biopanning of Zearalenone Mimotope by Phage Displayed Peptide Bank

目的:利用噬菌体肽库淘选玉米赤霉烯酮(zearalenone,ZEN)模拟表位。方法:以抗ZEN单克隆抗体为配体,利用噬菌体七肽库淘选ZEN的模拟表位,采用了逐轮减少抗体包被浓度,交换使用封阻液中蛋白质种类的方法,经三轮淘选后,随机挑取20个克隆测序并以ELISA方法及竞争ELISA实验,以确定阳性克隆。结果:获得10种序列,ELISA显示其中两种序列为阳性克隆,抑制率达90%以上,其氨基酸序列分别为DAVILLM,HHCHWWH。结论:噬菌体展示技术可淘选到ZEN的模拟表位,淘选到的噬菌体粒子可作为毒素的替代品建立免疫学检测方法。

Objective： To obtain mimotope of zearalenone （ZEN） by phage displayed peptide bank. Methods： The anti-ZEN monoclonal antibody was used as target for biopanning from a heptapeptide phage display bank. The concentration of antibody was reduced step by step, and different blocking buffer was alternated every round. After 3 rounds of biopanning, 20 clones were randomly selected for DNA sequencing, and assayed by ELISA and ZEN competition experiments to indentify the postive clones. Results： Ten sequences are obtained, two of which are proved to be positive. Inhibition percentages are up 90% and the amino acid sequences are DAVILLM, HHCHWWH respectively. Conclusion： The phage display technique can be successfully applied to panning the mimotope of ZEN.The aquired phages may be used as the surrogate of the toxin to establish immunoassay.