Division of Antibody Project, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi 470-1192, Japan.
Virology. 2010 Feb 20;397(2):322-30. doi: 10.1016/j.virol.2009.11.025. Epub 2009 Dec 6.
We tried to reveal the strain specificity of neutralizing mAbs against H3N2 influenza viruses in individuals. A large number of B lymphocytes of a pediatrician were collected by apheresis and two Ab libraries were constructed at 2004 and 2007 by using the phage-display technology. The libraries were screened against 12 different H3 strains of flu isolated between 1968 and 2004. Large numbers of clones that bound to the Ags were isolated and mAbs that specifically bound to H3 strain viruses were selected. Their binding activity to the 12 strains and neutralizing activity were studied by ELISA and focus reduction test, respectively. Furthermore, the binding activity to hemagglutinin (HA) was examined by Western blot. The majority of clones showing the neutralizing activity turned out to be anti-HA mAbs and could be divided into three major groups showing distinct strain specificity: 1968-1973, 1977-1993 and 1997-2003.
我们试图揭示针对 H3N2 流感病毒的中和单克隆抗体在个体中的菌株特异性。通过血细胞分离术收集了一位儿科医生的大量 B 淋巴细胞,并于 2004 年和 2007 年使用噬菌体展示技术构建了两个抗体文库。该文库针对 1968 年至 2004 年间分离出的 12 种不同的 H3 流感株进行了筛选。分离出大量与 Ag 结合的克隆,并选择了特异性结合 H3 株病毒的单克隆抗体。通过 ELISA 和空斑减少试验分别研究了它们对 12 个菌株的结合活性和中和活性。此外,通过 Western blot 检查了对血凝素 (HA) 的结合活性。表现出中和活性的大多数克隆是抗 HA 单克隆抗体,可分为显示出明显菌株特异性的三个主要组:1968-1973、1977-1993 和 1997-2003。