肺癌T7噬菌体文库中MUC1蛋白抗原模拟表位的筛选

Screening of MUC1 mimotope in T7 phage library of lung cancer

目的利用噬菌体表面展示技术,从肺癌T7噬菌体展示文库中筛选MUC1蛋白抗原的模拟表位。方法以MUC1单克隆抗体为靶分子,对肺癌T7噬菌体文库进行淘洗筛选,得到的阳性克隆进行测序并推导其氨基酸序列,对得到的阳性噬菌体克隆进行鉴定。结果经3轮淘选,得到20个阳性克隆。测序获得AAPDFRP、SAPDDRP 2种氨基酸序列,对MUC1单抗的抑制率均在50%以上,与肺癌血清结合特异性较高,此2种蛋白序列可模拟MUC1蛋白抗原表位。结论通过噬菌体展示技术成功筛选到MUC1蛋白的模拟表位序列XAPDXRP(X为任意氨基酸),对肺癌的早期诊断有一定的参考价值。

This study designed to screen MUC1 mimotope in T7 phage library of lung cancer by using phage surface display technology. T7 phage library of lung cancer was screened by biopanning with the anti-MUC1 monoclonal antibody acting as a molecular target, and the positive phage clones were sequenced and their amino acid sequences were derived for identification. After 3 rounds of biopanning, 20 positive clones were acquired. Two amino acid sequences, AAPDFRP and SAPDDRP, were obtained and the competitive inhibitory rate of MUC1 was more than 50%. Moreover, these positive clones showed higher serum binding specificity in lung cancer patients than in healthy people. These two protein sequences could be used to simulate the MUC1 protein epitopes. In conclusion, MUC1 mimotope has successfully screened by phage display technology, and its homologous sequence was XAPDXRP（X is any amino acid）, which may be a reference for the early diagnosis of lung cancer.