Dyson M R, Germaschewski V, Murray K
Institute of Cell and Molecular Biology, University of Edinburgh, UK.
Nucleic Acids Res. 1995 May 11;23(9):1531-5. doi: 10.1093/nar/23.9.1531.
Studies of interactions between filamentous fusion phage particles and protein or nucleic acid molecules have gained increasing importance with recent successes of screening techniques based upon random phage display libraries (biopanning). Since a number of different phage are usually obtained by biopanning, it is useful to compare quantitatively the binding affinities of individual phage for the substrate used for selection. A procedure is described for determination of relative dissociation constants (KdRel) between filamentous phage carrying peptide fusions to the coat protein gpIII and substrates in solution. This novel method is based on the measurement of phage titres. Phage selected from a random fusion phage library for binding to a monoclonal antibody or a viral structural protein exhibited KdRel values in the nanomolar and micromolar ranges for their respective substrates, thus validating the method over a wide range of binding affinities.
随着基于随机噬菌体展示文库（生物淘选）的筛选技术最近取得成功，丝状融合噬菌体颗粒与蛋白质或核酸分子之间相互作用的研究变得越来越重要。由于通常通过生物淘选获得多种不同的噬菌体，因此定量比较单个噬菌体对用于选择的底物的结合亲和力是有用的。本文描述了一种用于测定携带与外壳蛋白gpIII融合肽的丝状噬菌体与溶液中底物之间相对解离常数（KdRel）的方法。这种新方法基于噬菌体滴度的测量。从随机融合噬菌体文库中选择用于结合单克隆抗体或病毒结构蛋白的噬菌体，对其各自的底物表现出纳摩尔和微摩尔范围内的KdRel值，从而在广泛的结合亲和力范围内验证了该方法。
