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利用噬菌体随机肽库筛选抗TNF单抗表位的研究中,就抗体的选择、肽库富集的检测、筛选得到的表位多肽的验证及如何提高筛选的成功率等,进行了一些初步探索.实验结果表明,由识别线性抗原位点的抗体较容易筛选到噬菌体呈现表位,具有较强中和活性的抗体,因多识别空间构型抗原位点而增加筛选难度.NC膜斑点印迹、ELISA及DNA测序均可作为筛选富集的检测方法.用与天然抗原同源比较的方法及竞争性ELISA分析,可以帮助确定噬菌体呈现多肽是否是抗体表位
In the study of screening anti-TNF McAb epitopes by phage random peptide library, some preliminary studies were carried out on the selection of antibodies, the detection of peptide library enrichment, the verification of epitope peptides screened and how to improve the screening success rate . The experimental results show that it is easier to screen phage epitopes by antibodies recognizing linear antigenic sites, and antibodies with stronger neutralizing activity are more difficult to screen because of the recognition of the spatial configuration of antigenic sites. NC membrane blot, ELISA and DNA sequencing can be used as screening enrichment detection method. A method of homologous comparison with a native antigen and a competitive ELISA assay can help determine whether the phage-displayed polypeptide is an antibody epitope