Dybwad A, Bogen B, Natvig J B, Førre O, Sioud M
Institute of Immunology and Rheumatology, University of Oslo, Norway.
Clin Exp Immunol. 1995 Nov;102(2):438-42. doi: 10.1111/j.1365-2249.1995.tb03802.x.
We have examined the potential of isolating ligands for polyclonal antibodies from a nanopeptide phage library. The library was screened with a rabbit polyclonal antiserum raised against a synthetic peptide (ALWFRNHFVFGGGTKVT). Following screening, the positive phages were tested in an ELISA for their reactivity with the antiserum. Phages that showed positive reactivity with the antiserum compared with a normal rabbit serum were selected and their displayed peptides were determined. Among the 36 random positive clones, 31 clones carried the sequence AVFGGGTKL, PFFGGGSRA or APTGGSKRT that have a significant homology to the immunizing peptide. Five positive phages displayed the ATNIFIEGT sequence, which has no obvious linear homology with either the other selected peptides or with the peptide used for immunization. In contrast to the control peptide, the immunizing peptide inhibited binding of the antiserum to the peptide-displaying phages in a dose-dependent manner, thus demonstrating the specificity of the interaction. Furthermore, the rabbit B cell response to the peptide was found to be limited and focused on its C-terminal. Taken together, our data demonstrate the potential of random peptide phage libraries for defining epitopes for polyclonal antisera as well as for investigation of the nature of B cell responses to any given antigen.
我们研究了从纳米肽噬菌体文库中分离多克隆抗体配体的潜力。用针对合成肽（ALWFRNHFVFGGGTKVT）产生的兔多克隆抗血清筛选该文库。筛选后，在酶联免疫吸附测定（ELISA）中检测阳性噬菌体与抗血清的反应性。选择与正常兔血清相比与抗血清显示阳性反应性的噬菌体，并确定其展示的肽。在36个随机阳性克隆中，31个克隆携带与免疫肽具有显著同源性的序列AVFGGGTKL、PFFGGGSRA或APTGGSKRT。五个阳性噬菌体展示了ATNIFIEGT序列，该序列与其他选定的肽或用于免疫的肽均无明显的线性同源性。与对照肽相反，免疫肽以剂量依赖性方式抑制抗血清与展示肽的噬菌体的结合，从而证明了相互作用的特异性。此外，发现兔B细胞对该肽的反应有限且集中在其C末端。综上所述，我们的数据证明了随机肽噬菌体文库在确定多克隆抗血清表位以及研究B细胞对任何给定抗原反应性质方面的潜力。
