Slootstra J W, Puijk W C, Ligtvoet G J, Kuperus D, Schaaper W M, Meloen R H
Department of Molecular Recognition, Institute for Animal Science and Health (ID-DLO), Lelystad, The Netherlands.
J Mol Recognit. 1997 Sep-Oct;10(5):217-24.
Small diversity libraries, composed of 4550 synthetic dodecapeptides and 8000 synthetic tripeptides, have been used to identify sequences homologous to small linear and non-linear parts of epitopes. Here we report that synthetic peptides identified through alignment of dodecapeptides and tripeptides derived from these small libraries have, in direct ELISA and/or competitive ELISA, activities similar to that of peptides covering the native epitope and similar to that of peptides derived from large expression libraries composed of 10(6)-10(7) random peptides. This result was obtained with the monoclonal antibodies 6A.A6 and M2. Mab 6A.A6 binds the transmissible gastroenteritis virus (TGEV) and mAb M2 binds the FLAG-peptide, an affinity tag. It was also found that the antibody binding activity of peptides, derived from small or large libraries, can strongly depend on the way in which the peptide is presented to the antibody, i.e. high antibody titers were obtained when these peptides were synthesized on pins or coated onto microtiter plates, whereas low IC50s were obtained with these peptides in solution. We postulate that small peptide libraries may be a powerful tool to quickly identify new peptides that can be used as sensitive markers for mAbs of interest.
由4550种合成十二肽和8000种合成三肽组成的小型多样性文库已被用于鉴定与表位的小线性和非线性部分同源的序列。在此我们报告，通过比对源自这些小型文库的十二肽和三肽而鉴定出的合成肽，在直接ELISA和/或竞争性ELISA中，其活性与覆盖天然表位的肽相似，且与源自由10(6)-10(7)个随机肽组成的大型表达文库的肽相似。该结果是用单克隆抗体6A.A6和M2获得的。单克隆抗体6A.A6结合传染性胃肠炎病毒（TGEV），单克隆抗体M2结合FLAG肽（一种亲和标签）。还发现，源自小型或大型文库的肽的抗体结合活性可能强烈取决于肽呈递给抗体的方式，即当这些肽在针上合成或包被在微量滴定板上时可获得高抗体滴度，而这些肽在溶液中时则获得低IC50值。我们推测小型肽文库可能是快速鉴定可作为感兴趣的单克隆抗体的敏感标志物的新肽的有力工具。
