Dueñas M, Borrebaeck C A
Department of Immunotechnology, Lund University, Sweden.
Biotechnology (N Y). 1994 Oct;12(10):999-1002. doi: 10.1038/nbt1094-999.
The immune response generates a tremendous array of antibody specificities by VDJ-gene rearrangements. A similar diversity can be obtained by expressing entire V-gene repertoires on the surface of filamentous bacteriophages creating large antibody libraries. Here we describe how the clonal selection mechanisms of the humoral immune response can also be mimicked in the phage display system by linking antigen-recognition and phage replication. We have achieved this by displaying antibody libraries on engineered, non-infectious phage with gene 3 deletions. Individual, antigen-specific phage are made replication competent by allowing a fusion protein, consisting of the antigen and phage coat protein 3, to bind the displayed antibody fragment. This fusion protein bridges the phage and F-pili of the bacteria and allows infection to be initiated and the phage to be clonally amplified with specific enrichment factors of approximately 10(10) after only two rounds.
免疫反应通过VDJ基因重排产生大量抗体特异性。通过在丝状噬菌体表面表达完整的V基因库来创建大型抗体文库，也可以获得类似的多样性。在这里，我们描述了如何通过将抗原识别与噬菌体复制联系起来，在噬菌体展示系统中模拟体液免疫反应的克隆选择机制。我们通过在具有基因3缺失的工程化非感染性噬菌体上展示抗体文库实现了这一点。通过允许由抗原和噬菌体外壳蛋白3组成的融合蛋白结合展示的抗体片段，使单个抗原特异性噬菌体具有复制能力。这种融合蛋白连接噬菌体和细菌的F菌毛，并允许启动感染，并且仅经过两轮后，噬菌体就可以以约10(10)的特异性富集因子进行克隆扩增。
