黄曲霉毒素M_1模拟表位的筛选和鉴定

Screen and Identification of AFM_1 Mimic Epitope

目的:从噬菌体表面7肽库中筛选出黄曲霉毒素M1抗原的模拟表位。方法与结果:经过4轮淘选,筛选获得了4个与抗AFM1单克隆抗体具有较高亲和性的阳性噬菌体克隆,编号为A1、A2、A3和A4。间接竞争抑制实验结果表明:当噬菌体滴度为2.5×1010CFU时,噬菌体A1、A2、A3和A4对AFM1与抗AFM1单克隆抗体结合的抑制率分别为41.8%、40.6%、36.7%和37.6%。测定其核酸序列,其中A1、A2为同一克隆,氨基酸序列为LTSFPRH;A3、A4为同一克隆,氨基酸序列为MAPSSWR。结论:从噬菌体7肽库中筛选到2个AFM1的理想模拟表位。

Objective： To screen AFM1 mimic epitope by using phage display peptide library technology. Methods and Results： After three rounds of biopanning, 4 positive phage clones designated as A1, A2, A3 and A4 were pick out and found that all of them could block the binding of AFM1 to antigen when the pfu of phage was 2.5 × 10^10. The inhibition rates were 41.8%, 40.6%, 36.7% and 37.6% respectively. Results of DNA analysis showed that A1 and A2 belong to the same clone and their identical amino acid sequence is LTSFPRH; A3 and A4 belong to the same clone and their iolentical amino acid sequence is MAPSSWR. Conclusions： 2 AFM1 mimic epitopes were obtained from phage display peptide library.