Phage display of cDNA repertoires:: the pVI display system and its applications for the selection of immunogenic ligands
Phage display of cDNA repertoires:: the pVI display system and its applications for the selection of immunogenic ligands
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DOI：
10.1016/s0022-1759(99)00139-8
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发表时间：
1999-12-10
影响因子：
2.2
通讯作者：
Hoogenboom, HR
中科院分区：
文献类型：
--
作者：
Hufton, SE;Moerkerk, PT;Hoogenboom, HR
据报道，在固定的靶标上选择噬菌体展示的cDNA库是快速识别相互作用伙伴的一种有效方法。然而，到目前为止，只有几个成功的申请被报道。本文就噬菌体展示和筛选文库的研究现状作一综述。作为一个例子，我们报道了一组基于与丝状噬菌体的细小噬菌体外壳蛋白6(PVI)融合而构建的适合于cDNA展示的噬菌体展示载体。我们通过展示人免疫球蛋白C(H)3结构域和大肠杆菌碱性磷酸酶(PhoA)基因对这些载体进行了评价。免疫球蛋白G和PhoA的C(H)3结构域都显示在PVI上，PhoA也具有酶活性。我们已经在这些载体中构建了原代大肠肿瘤基因编码库，并对抗β2微球蛋白的单抗(β2M)和抗人免疫球蛋白的多克隆抗体血清进行了筛选。在这两种情况下，相关的配体都是从噬菌体展示的cDNA库中恢复的。这些载体可用于用患者多克隆血清筛选噬菌体展示文库。这将使人们能够在癌症、病毒/细菌感染或自身免疫性疾病等疾病中识别抗原性cDNA产物。此外，通过与其他特定生物分子的选择，该展示系统可能有助于识别功能基因组学中的相互作用伙伴。(C)1999 Elsevier Science B.V.保留所有权利。
The selection of phage displayed cDNA repertoires on an immobilised target has been reported to be an efficient way to rapidly identify interacting partners. To date, however, only a few successful applications have been reported. Here, we present a review of the current status of the display and selection of cDNA libraries using phage. As an example, we report the construction of a set of phage display vectors suitable for cDNA display based on fusion to the minor bacteriophage coat protein 6 (pVI) of filamentous phage. We have evaluated these vectors through the display of the C(H)3 domain of human IgG and of the Escherichia coli alkaline phosphatase (PhoA) gene. Both the C(H)3 domain of IgG and PhoA are shown to be displayed on pVI, and PhoA is also shown to be enzymatically active. We have constructed primary colorectal tumor cDNA repertoires in these vectors and performed selections on both a monoclonal antibody to beta 2 microglobulin (beta 2M) and polyclonal antibody sera to human IgG. In both cases, relevant ligands were recovered from the phage displayed cDNA repertoire. These vectors may be used for selection of phage displayed cDNA libraries with polyclonal sera from patients. This will allow the identifying antigenic cDNA products in such diseases as cancer, viral/bacterial infections or autoimmune disease. Furthermore, by selections with other specific biomolecules, this display system may aid the identification of interacting partners in functional genomics. (C) 1999 Elsevier Science B.V. All rights reserved.