Fine epitope mapping of monoclonal antibodies against hemagglutinin of a highly pathogenic H5N1 influenza virus using yeast surface display.
Fine epitope mapping of monoclonal antibodies against hemagglutinin of a highly pathogenic H5N1 influenza virus using yeast surface display.
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DOI：
10.1016/j.bbrc.2011.04.139
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发表时间：
2011-06-03
影响因子：
3.1
通讯作者：
Marasco, Wayne A.
中科院分区：
文献类型：
--
作者：
Han, Thomas;Sui, Jianhua;Bennett, Andrew S.;Liddington, Robert C.;Donis, Ruben O.;Zhu, Quan;Marasco, Wayne A.
► 我们使用强大的酵母展示系统绘制了流感 HA 抗体图谱。 ► 全长 H5 亚型 HA 在酵母上以正确的构象表达。 ► 使用酵母展示的 HA1 突变体文库来精细绘制抗 HA mAb 表位。 ► 强效中和性 NR2728 mAb 与受体结合位点相互作用。 ► H5-2A mAb 专门定位到仅在 H5 HA 上发现的高度保守表位。高致病性 H5N1 禽流感病毒构成了令人衰弱的大流行威胁。因此，了解抗体介导的病毒抑制和中和逃逸机制至关重要。在这里，展示了一种强大的酵母展示系统，用于病毒表面血凝素 (HA) 特异性抗体的精细表位作图。全长 H5 亚型 HA (HA0) 在酵母表面以正确折叠的构象表达，这是通过一组广泛表征的中和人单克隆抗体 (mAb) 的结合来确定的。这些 mAb 靶向甲型流感 HA 的构象依赖性表位，该表位在 H5 进化枝和 1 组血清型中高度保守。通过在酵母上分别展示 HA1 和 HA2 亚基，对两种抗 H5 单克隆抗体 NR2728 和 H5-2A 进行域作图，将其表位定位于 HA1。通过使用酵母展示的 HA1 突变体文库并在事先不了解潜在接触残基的情况下选择结合丧失，对这些抗 H5 mAb 表位进行了进一步精细定位。通过将影响结合的关键突变残基覆盖到 HA 晶体结构上，发现 NR2728 mAb 与受体结合位点 (RBS) 处完全表面暴露的连续残基相互作用，从而深入了解其有效抑制病毒结合的机制。非中和性 H5-2A mAb 类似地被映射到三聚体 HA 界面环上高度保守的 H5 菌株特异性但不易接近的位置。这些数据进一步增强了我们研究针对自然和实验性流感感染和疫苗的 HA 抗原性、表位多样性和可及性的工具箱。
► We mapped influenza HA antibodies using a robust yeast display system. ► The full-length H5 subtype HA was expressed on yeast in the proper conformation. ► Mutant library of yeast-displayed HA1 was used to fine map anti-HA mAbs epitopes. ► The potent neutralizing NR2728 mAb interacts with the receptor binding site. ► H5-2A mAb mapped specifically to a highly conserved epitope only found on H5 HA. Highly pathogenic H5N1 avian influenza viruses pose a debilitating pandemic threat. Thus, understanding mechanisms of antibody-mediated viral inhibition and neutralization escape is critical. Here, a robust yeast display system for fine epitope mapping of viral surface hemagglutinin (HA)-specific antibodies is demonstrated. The full-length H5 subtype HA (HA0) was expressed on the yeast surface in a correctly folded conformation, determined by binding of a panel of extensively characterized neutralizing human monoclonal antibodies (mAbs). These mAbs target conformationally-dependent epitopes of influenza A HA, which are highly conserved across H5 clades and group 1 serotypes. By separately displaying HA1 and HA2 subunits on yeast, domain mapping of two anti-H5 mAbs, NR2728 and H5-2A, localized their epitopes to HA1. These anti-H5 mAb epitopes were further fine mapped by using a library of yeast-displayed HA1 mutants and selecting for loss of binding without prior knowledge of potential contact residues. By overlaying key mutant residues that impacted binding onto a crystal structure of HA, the NR2728 mAb was found to interact with a fully surface-exposed contiguous patch of residues at the receptor binding site (RBS), giving insight into the mechanism underlying its potent inhibition of virus binding. The non-neutralizing H5-2A mAb was similarly mapped to a highly conserved H5 strain-specific but poorly accessible location on a loop at the trimer HA interface. These data further augment our toolchest for studying HA antigenicity, epitope diversity and accessibility in response to natural and experimental influenza infection and vaccines.
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开发具有针对西尼罗河病毒的治疗潜力的人源化单克隆抗体。
影响因子：
82.9
作者：
Oliphant, Theodore;Engle, Michael;Nybakken, Grant E;Doane, Chris;Johnson, Syd;Huang, Ling;Gorlatov, Sergey;Mehlhop, Erin;Marri, Anantha;Chung, Kyung Min;Ebel, Gregory D;Kramer, Laura D;Fremont, Daved H;Diamond, Michael S
通讯作者：
Diamond, Michael S
DOI：
10.1128/cdli.5.1.114-117.1998
发表时间：
1998-01-01
期刊：
CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY
影响因子：
--
作者：
Benne, CA;Kroon, FP;De Jong, JC
通讯作者：
De Jong, JC
影响因子：
5.6
作者：
Levy, R.;Forsyth, C. M.;Marks, J. D.
通讯作者：
Marks, J. D.
DOI：
10.1038/nri2054
发表时间：
2007-04
期刊：
Nature reviews. Immunology
影响因子：
--
作者：
Subbarao K;Joseph T
通讯作者：
Joseph T
影响因子：
2.2
作者：
Cochran, JR;Kim, YS;Wittrup, KD
通讯作者：
Wittrup, KD