MAKING ANTIBODY FRAGMENTS USING PHAGE DISPLAY LIBRARIES
MAKING ANTIBODY FRAGMENTS USING PHAGE DISPLAY LIBRARIES
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DOI：
10.1038/352624a0
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发表时间：
1991-08-15
期刊：
影响因子：
64.8
通讯作者：
WINTER, G
中科院分区：
文献类型：
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作者：
CLACKSON, T;HOOGENBOOM, HR;WINTER, G
为了绕过杂交瘤技术和动物免疫，我们正试图通过模仿免疫选择的特征在细菌中制造抗体。最近，我们使用Fd噬菌体2展示了融合到次要外壳蛋白3，4的抗体片段，从而允许用抗原3丰富噬菌体。利用重排重链(VH)和kappa轻链(V-kappa)轻链5-8的随机组合文库，我们现在已经在Fd噬菌体表面展示了不同的抗体片段文库。单次通过半抗原亲和柱后，检测到具有一系列Phox结合活性的Fd噬菌体，至少有一个具有高亲和力(解离常数，K(D)=10(-8)M)。第二次传球以牺牲弱者为代价，为强者增加了财富。粘合剂是由V基因编码的，类似于在抗PHOX杂交瘤中发现的，但在混杂组合中(在几个不同的伴侣中发现相同的V基因)。通过将混杂的VH或V-kappa基因与不同的伙伴谱系相结合来创建分层文库，我们获得了更多具有强烈结合活性的配对。噬菌体展示提供了从V基因库制造抗体、改变V结构域配对和选择亲和力良好的抗体的新方法。
To by-pass hybridoma technology and animal immunization, we are trying to build antibodies in bacteria by mimicking features of immune selection1. Recently we used fd phage2 to display antibody fragments fused to a minor coat protein3,4, allowing enrichment of phage with antigen3. Using a random combinatorial library of the rearranged heavy (VH) and kappa (V-kappa) light chains5-8 from mice immune to the hapten 2-phenyloxazol-5-one (phOx), we have now displayed diverse libraries of antibody fragments on the surface of fd phage. After a single pass over a hapten affinity column, fd phage with a range of phOx binding activities were detected, at least one with high affinity (dissociation constant, K(d) = 10(-8) M). A second pass enriched for the strong binders at the expense of the weak. The binders were encoded by V genes similar to those found in anti-phOx hybridomas but in promiscuous combinations (where the same V gene is found with several different partners). By combining a promiscuous VH or V-kappa gene with diverse repertoires of partners to create hierarchical libraries, we elicited many more pairings with strong binding activities. Phage display offers new ways of making antibodies from V-gene libraries, altering V-domain pairings and selecting for antibodies with good affinities.