Bugli Francesca, Graffeo Rosalia, Pescatori Mario, Paroni Sterbini Francesco, Torelli Riccardo, Masucci Luca, Manzara Stefania, Fadda Giovanni
Institute of Microbiology, Catholic University, Rome, Italy.
New Microbiol. 2009 Oct;32(4):341-9.
Evidence from clinical and experimental studies indicates that hepatitis C virus E2 glycoprotein (HCV/E2) represents a major target antigen involved in the containment and resolution of naturally occurring HCV infection. Antibody phage display allows the molecular cloning of cDNA sequences encoding antibody fragments specific to a wide range of diverse antigens. These antibodies may be produced in bacteria as Fab or converted into full length IgG. The latter have a higher serum half life and display Fc encoded function. Using a library prepared from an HCV-infected individual, we selected a panel of Fab fragments for binding to invariant epitopes of the E2 glycoprotein. This work describes a technique used to convert the selected Fab fragments into full length IgG and to express these antibodies in eukaryotic cells. All the recombinant antibodies retained the binding specificity of the parental Fab showing an increase in apparent relative affinity for E2.
临床和实验研究证据表明，丙型肝炎病毒E2糖蛋白（HCV/E2）是参与自然发生的HCV感染控制和消退的主要靶抗原。抗体噬菌体展示技术可实现对编码针对多种不同抗原的特异性抗体片段的cDNA序列进行分子克隆。这些抗体可以在细菌中作为Fab片段产生，或转化为全长IgG。后者具有更长的血清半衰期并表现出Fc编码的功能。利用从一名HCV感染者制备的文库，我们筛选出了一组与E2糖蛋白恒定表位结合的Fab片段。本文描述了一种将筛选出的Fab片段转化为全长IgG并在真核细胞中表达这些抗体的技术。所有重组抗体都保留了亲本Fab的结合特异性，对E2的表观相对亲和力有所增加。
