Plaisant P, Burioni R, Manzin A, Solforosi L, Candela M, Gabrielli A, Fadda G, Clementi M
Istituto di Microbiologia, Facoltà di Medicina, Università Cattolica del Sacro Cuore, Roma, Italy.
Res Virol. 1997 Mar-Apr;148(2):165-9. doi: 10.1016/s0923-2516(97)89904-9.
Molecular cloning of the antibody repertoire in phage display combinatorial vectors is a powerful method enabling the dissection of the immunoresponse against a given pathogen. In this paper we describe the construction of a combinatorial library displayed on phage surface, containing the antibody repertoire of a patient with high serological response against hepatitis C virus (HCV) antigens. Following selection of the library against solid-phase-bound antigen, sixteen human antibody Fab fragments able to bind to HCV-specific antigens were generated and studied for binding characteristics. The majority of them appeared to have specificity for the HCV c33 peptide. All the clones reacting with the c33 peptide shared the same heavy-chain CDR3 sequence. This is the first report of molecular cloning in a combinatorial phage display vector of the antibody repertoire of an anti-HCV-positive patient.
在噬菌体展示组合载体中对抗体库进行分子克隆是一种强大的方法，能够剖析针对特定病原体的免疫反应。在本文中，我们描述了构建一个展示在噬菌体表面的组合文库，该文库包含一名对丙型肝炎病毒（HCV）抗原具有高血清学反应的患者的抗体库。在用固相结合抗原筛选该文库后，产生了16个能够结合HCV特异性抗原的人抗体Fab片段，并对其结合特性进行了研究。它们中的大多数似乎对HCV c33肽具有特异性。所有与c33肽反应的克隆都共享相同的重链CDR3序列。这是关于抗HCV阳性患者抗体库在组合噬菌体展示载体中的分子克隆的首次报道。
