Kogot Joshua M, Pennington Joseph M, Sarkes Deborah A, Kingery David A, Pellegrino Paul M, Stratis-Cullum Dimitra N
US Army Research Laboratory; Sensors and Electron Devices, Adelphi, MD, USA.
J Mol Recognit. 2014 Dec;27(12):739-45. doi: 10.1002/jmr.2400.
Bacterial peptide display libraries enable the rapid and efficient selection of peptides that have high affinity and selectivity toward their targets. Using a 15-mer random library on the outer surface of Escherichia coli (E.coli), high-affinity peptides were selected against a staphylococcal enterotoxin B (SEB) protein after four rounds of biopanning. On-cell screening analysis of affinity and specificity were measured by flow cytometry and directly compared to the synthetic peptide, off-cell, using peptide-ELISA. DNA sequencing of the positive clones after four rounds of microfluidic magnetic sorting (MMS) revealed a common consensus sequence of (S/T)CH(Y/F)W for the SEB-binding peptides R338, R418, and R445. The consensus sequence in these bacterial display peptides has similar amino acid characteristics with SEB peptide sequences isolated from phage display. The Kd measured by peptide-ELISA off-cell was 2.4 nM for R418 and 3.0 nM for R445. The bacterial peptide display methodology using the semiautomated MMS resulted in the discovery of selective peptides with affinity for a food safety and defense threat. Published 2014. This article is a U.S. Government work and is in the public domain in the USA.
细菌肽展示文库能够快速有效地筛选出对其靶标具有高亲和力和选择性的肽。利用大肠杆菌（E.coli）外表面的一个15聚体随机文库，经过四轮生物淘选，筛选出了针对葡萄球菌肠毒素B（SEB）蛋白的高亲和力肽。通过流式细胞术对亲和力和特异性进行细胞表面筛选分析，并使用肽酶联免疫吸附测定法（peptide-ELISA）将其与合成肽（细胞外）进行直接比较。经过四轮微流控磁分选（MMS）后，对阳性克隆进行DNA测序，结果显示SEB结合肽R338、R418和R445的共有一致序列为（S/T）CH（Y/F）W。这些细菌展示肽中的共有序列与从噬菌体展示中分离出的SEB肽序列具有相似的氨基酸特征。通过细胞外肽酶联免疫吸附测定法测得，R418的解离常数（Kd）为2.4 nM，R445的解离常数为3.0 nM。使用半自动微流控磁分选的细菌肽展示方法，发现了对食品安全和防御威胁具有亲和力的选择性肽。2014年发表。本文为美国政府作品，在美国属于公共领域。
