Probing the basis of antibody reactivity with a panel of constrained peptide libraries displayed by filamentous phage
Probing the basis of antibody reactivity with a panel of constrained peptide libraries displayed by filamentous phage
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DOI：
10.1006/jmbi.1996.0284
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发表时间：
1996-05-24
影响因子：
5.6
通讯作者：
Scott, JK
中科院分区：
文献类型：
--
作者：
Bonnycastle, LLC;Mehroke, JS;Scott, JK
肽与抗体结合的结构要求可以通过用具有不同限制的多种肽进行探测来阐明。为此，我们构建并筛选了一组丝状噬菌体展示的肽库。大多数文库中的肽有可能受到固定Cys残基的限制，这些残基被放置在不同长度的随机氨基酸序列中的不同位点。当综合起来时，通过用给定抗体筛选组而获得的结合数据允许人们确定促进结合的限制类型，以及对结合至关重要的残基。我们描述了 11 个 pVIII 展示的肽文库的构建，其大小范围从 1.5 亿到 100 亿个克隆。使用许多针对肽、蛋白质和碳水化合物的多克隆和单克隆抗体对文库进行了筛选。针对肽、折叠蛋白质上的线性表位以及令人惊讶的碳水化合物产生的抗体总是发现与肽的交叉反应性，而针对蛋白质上的不连续表位的抗体则很少被发现。这些结果的含义根据与肽交叉反应性的结构基础进行了讨论。 (C) 1996 学术出版社有限公司
The structural requirements for peptide binding to an antibody may be elucidated by probing it with a variety of peptides having different constraints. To this end, we have constructed and screened a panel of peptide libraries displayed by filamentous bacteriophage. The peptides in most of the libraries have the potential for constraint by fixed Cys residues, which have been placed at different sites within a randomized amino acid sequence of varying length. When taken together, the binding data obtained from screening the panel with a given antibody allow one to determine the types of constraints that promote binding, as well as the residues that are critical for binding. We describe the construction of 11, pVIII-displayed, peptide libraries, whose sizes range from 150 million to 10 billion clones. The libraries were screened with a number of polyclonal and monoclonal antibodies against peptides, proteins and carbohydrates. Cross-reactivity with peptides was always found for antibodies produced against peptides, linear epitopes on folded proteins and, surprisingly, carbohydrates, whereas antibodies against discontinuous epitopes on proteins were found less frequently The implications of these results are discussed in terms of the structural basis for cross-reactivity with peptides. (C) 1996 Academic Press Limited