半抗原 DNP 的肽模拟物：抗 DNP 单克隆抗体对噬菌体展示模拟物的亲和力选择的影响。

Peptide mimics of hapten DNP: the effect of affinity of anti-DNP monoclonal antibodies for the selection of phage-displayed mimotopes.

机构信息

Department of Pathology and Infectious Diseases, Royal Veterinary College, University of London, Royal College Street, London NW1 0TU, UK.

出版信息

J Immunol Methods. 2011 Mar 7;366(1-2):36-42. doi: 10.1016/j.jim.2011.01.004. Epub 2011 Jan 22.

DOI:10.1016/j.jim.2011.01.004

PMID:21262229

Abstract

Biopanning of two linear (6- and 15-mer) and two constrained (10- and 17-mer) phage-displayed peptide libraries with two anti-DNP monoclonal antibodies (mAbs) selected seven unique peptide sequences using only the low affinity anti-DNP monoclonal antibody. The selected peptides contained two of 6, one of 10, two of 15 and two of 17 amino acids in length. They were all rich in hydrophobic residues. Both 15-mer peptides had antigenic regions of eight amino acids as revealed by a spot scan assay. Two of the 17-mer and one of the 10-mer peptides displayed on phage competed with free DNP for the low affinity anti-DNP mAb. These findings highlight (i) the selective power of phage displayed peptide libraries to identify peptides that mimic the shape of a small hapten molecule such as DNP, (ii) the possible preferential bias of phage libraries towards low affinity antibodies, (iii) the importance of using a panel of phage libraries for selecting peptide mimics.

摘要

用两种抗 DNP 单克隆抗体（mAb）对两个线性（6-和 15-mer）和两个受限（10-和 17- mer）噬菌体展示肽文库进行生物淘选，仅使用低亲和力抗 DNP mAb 就选择了七个独特的肽序列。所选肽段的长度分别为 6 个氨基酸、10 个氨基酸、15 个氨基酸和 17 个氨基酸。它们都富含疏水性残基。斑点扫描分析显示，两个 15 肽和一个 10 肽均具有 8 个氨基酸的抗原区域。两个 17 肽和一个 10 肽在噬菌体上显示的肽与游离 DNP 竞争低亲和力抗 DNP mAb。这些发现突出了：（i）噬菌体展示肽文库具有识别类似 DNP 等小分子半抗原形状的肽的选择性；（ii）噬菌体文库可能对低亲和力抗体有偏好性；（iii）使用多组噬菌体文库选择肽模拟物的重要性。