Desogus Alessandra, Burioni Roberto, Ingianni Angela, Bugli Francesca, Pompei Raffaello, Fadda Giovanni
Sezione di Microbiologia Applicata, Dipartimento di Scienze e Tecnologie Biomediche, Università di Cagliari, Italy.
Clin Diagn Lab Immunol. 2003 Jul;10(4):680-5. doi: 10.1128/cdli.10.4.680-685.2003.
A human recombinant monoclonal Fab fragment that specifically recognizes all the influenza A virus strains tested was produced in transformed Escherichia coli using the phage display technique. No strain of influenza B virus reacted with it. It was purified after four cycles of panning and by a single passage through an immunoaffinity column. About 1 mg of pure monoclonal antibody was obtained from 1 liter of culture medium in 3 working days. The Fab fragment reacted with a viral 27-kDa protein, which could reasonably be a matrix protein. Indirect immunofluorescence tests performed on virus-infected MDCK cells showed that this Fab fragment was at least equally efficient as other commercial monoclonal antibody-based systems in detecting influenza A viral infections. The potential advantages of human recombinant Fabs on murine monoclonal antibodies are discussed.
利用噬菌体展示技术在转化的大肠杆菌中制备了一种特异性识别所有测试甲型流感病毒株的人重组单克隆Fab片段。没有乙型流感病毒株与它发生反应。经过四轮淘选并通过免疫亲和柱单次过柱后进行纯化。在3个工作日内从1升培养基中获得了约1毫克的纯单克隆抗体。该Fab片段与一种病毒27 kDa蛋白发生反应,该蛋白合理推测可能是一种基质蛋白。在病毒感染的MDCK细胞上进行的间接免疫荧光试验表明,该Fab片段在检测甲型流感病毒感染方面至少与其他基于单克隆抗体的商业系统同样有效。文中讨论了人重组Fab相对于鼠单克隆抗体的潜在优势。