Influenza PR8 HA-specific Fab fragments produced by phage display methods
Influenza PR8 HA-specific Fab fragments produced by phage display methods
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DOI：
10.1016/j.bbrc.2007.11.135
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发表时间：
2008-02-08
影响因子：
3.1
通讯作者：
Fujita-Yamaguchi, Yoko
中科院分区：
文献类型：
--
作者：
Asanuma, Hideki;Matsumoto-Takasaki, Ayano;Fujita-Yamaguchi, Yoko
以流感病毒a /Puerto Rico/8/34 （PR8; H1N1）纯化的HA为抗原，从噬菌体展示文库中分离到抗流感血凝素（HA） fab。选择4个对PR8 HA的结合信号比对照高25 - 50倍的Fab克隆进行进一步分析，并与抗PR8单克隆抗体（mAb）进行比较。所有四种fab和mAb在非还原条件下识别PR8 HA，但很少与还原的PR8 HA结合。Fab1和mAb对流感病毒感染MDCK细胞的抑制作用呈剂量依赖性，而Fab3和4仅表现出部分抑制作用。此外，Fab1克隆和单抗与A/Peking/262/95 （A/Peking; H1N1）病毒具有交叉反应性。mAb对两种流感病毒株的抑制作用比Fab1更强，这可能是由于其对抗原的亲和力更高。事实上，SPR分析显示，Fab1和mAb的K-D分别为1.5 × 10（-8）和3.2 × 10(-9) M。这些结果强烈提示，噬菌体文库衍生的fab可以很容易地制备出来，并且这种对流感感染具有抑制作用的ha特异性fab可能用于治疗流感患者。(c) 2007爱思唯尔公司版权所有。
Anti-influenza hemagglutinin (HA) Fabs were isolated from a phage display library using purified HA of influenza virus A/Puerto Rico/8/34 (PR8; H1N1) as an antigen. Four Fab clones displaying a 25 - 50-fold higher binding signal to PR8 HA than the control were selected for further analysis and comparison with anti-PR8 monoclonal antibody (mAb). All four Fabs and mAb recognized the PR8 HA under non-reducing conditions but rarely bound to reduced PR8 HA. Inhibition of influenza virus infection on MDCK cells was observed with Fab1 and mAb in a dose-dependent manner while Fab3 and 4 exhibited only a partial inhibitory effect. Moreover, Fab1 clone and mAb exhibited cross-reactivity with the A/Peking/262/95 (A/Peking; H1N1) strain. The inhibitory effects of mAb on both influenza strains were more potent than Fab1, which is likely attributed to its higher affinity for the antigen. SPR analyses, in fact, revealed that Fab1 and mAb have K-D of 1.5 x 10(-8) and 3.2 x 10(-9) M, respectively. These results strongly suggest that phage library-derived Fabs can be readily prepared and that such HA-specific Fabs with inhibitory action on influenza infection may be used to treat influenza patients. (c) 2007 Elsevier Inc. All rights reserved.