Scott J K, Smith G P
Division of Biological Sciences, University of Missouri, Columbia 65211.
Science. 1990 Jul 27;249(4967):386-90. doi: 10.1126/science.1696028.
Tens of millions of short peptides can be easily surveyed for tight binding to an antibody, receptor or other binding protein using an "epitope library." The library is a vast mixture of filamentous phage clones, each displaying one peptide sequence on the virion surface. The survey is accomplished by using the binding protein to affinity-purify phage that display tight-binding peptides and propagating the purified phage in Escherichia coli. The amino acid sequences of the peptides displayed on the phage are then determined by sequencing the corresponding coding region in the viral DNA's. Potential applications of the epitope library include investigation of the specificity of antibodies and discovery of mimetic drug candidates.
使用“表位文库”,可以轻松筛选数以千万计的短肽,以寻找与抗体、受体或其他结合蛋白的紧密结合。该文库是丝状噬菌体克隆的大量混合物,每个克隆在病毒体表面展示一个肽序列。通过使用结合蛋白对展示紧密结合肽的噬菌体进行亲和纯化,并在大肠杆菌中繁殖纯化后的噬菌体来完成筛选。然后,通过对病毒DNA中相应编码区域进行测序,确定噬菌体上展示的肽的氨基酸序列。表位文库的潜在应用包括研究抗体的特异性和发现模拟药物候选物。