Slootstra J W, Puijk W C, Ligtvoet G J, Langeveld J P, Meloen R H
Department of Molecular Recognition, Institute for Animal Science and Health (ID-DLO), Lelystad, The Netherlands.
Mol Divers. 1996 Feb;1(2):87-96. doi: 10.1007/BF01721323.
Two small random peptide libraries, one composed of 4550 dodecapeptides and one of 8000 tripeptides, were synthesized in newly developed credit-card format miniPEPSCAN cards (miniPEPSCAN libraries). Each peptide was synthesized in a discrete well (455 peptides/card). The two miniPEPSCAN libraries were screened with three different monoclonal antibodies (Mabs). Two other random peptide libraries, expressed on the wall of bacteria (recombinant libraries) and composed of 10(7) hexa- and octapeptides, were screened with the same three Mabs. The aim of this study was to compare the amino acid sequence of peptides selected from small and large pools of random peptides and, in this way, investigate the potential of small random peptide libraries. The screening of the two miniPEPSCAN libraries resulted in the identification of a surprisingly large number of antibody-binding peptides, while the screening of the large recombinant libraries, using the same Mabs, resulted in the identification of only a small number of peptides. The large number of peptides derived from the small random peptide libraries allowed the determination of consensus sequences. These consensus sequences could be related to small linear and nonlinear parts of the respective epitopes. The small number of peptides derived from the large random peptide libraries could only be related to linear epitopes that were previously mapped using small libraries of overlapping peptides covering the antigenic protein. Thus, with respect to the cost and speed of identifying peptides that resemble linear and nonlinear parts of epitopes, small diversity libraries based on synthetic peptides appear to be superior to large diversity libraries based on expression systems.
合成了两个小型随机肽库，一个由4550个十二肽组成，另一个由8000个三肽组成，它们以新开发的信用卡格式微型PEPSCAN卡（微型PEPSCAN库）形式合成。每个肽在一个独立的孔中合成（每张卡455个肽）。用三种不同的单克隆抗体（Mab）筛选这两个微型PEPSCAN库。另外两个在细菌壁上表达（重组库）、由10⁷个六肽和八肽组成的随机肽库，也用同样的三种单克隆抗体进行筛选。本研究的目的是比较从小型和大型随机肽库中筛选出的肽的氨基酸序列，从而研究小型随机肽库的潜力。对两个微型PEPSCAN库的筛选鉴定出了数量惊人的抗体结合肽，而用同样的单克隆抗体筛选大型重组库时，只鉴定出了少数肽。从小型随机肽库中获得的大量肽使得能够确定共有序列。这些共有序列可能与各自表位的小线性和非线性部分相关。从大型随机肽库中获得的少数肽只能与先前使用覆盖抗原蛋白的重叠肽小型库定位的线性表位相关。因此，就鉴定类似于表位线性和非线性部分的肽的成本和速度而言，基于合成肽的小型多样性库似乎优于基于表达系统的大型多样性库。
