基于生物传感器的针对甲型流感病毒血凝素和神经氨酸酶的抗体表位作图。
Biosensor-based epitope mapping of antibodies targeting the hemagglutinin and neuraminidase of influenza A virus.
机构信息
Influenza Division, National Center for Immunization and Respiratory Disease, Centers for Disease Control and Prevention, Atlanta, GA, USA.
Influenza Division, National Center for Immunization and Respiratory Disease, Centers for Disease Control and Prevention, Atlanta, GA, USA; CNI Advantage, LLC, Norman, OK, USA.
出版信息
J Immunol Methods. 2018 Oct;461:23-29. doi: 10.1016/j.jim.2018.07.007. Epub 2018 Jul 24.
Abstract
Characterization of the epitopes on antigen recognized by monoclonal antibodies (mAb) is useful for the development of therapeutic antibodies, diagnostic tools, and vaccines. Epitope mapping also provides functional information for sequence-based repertoire analysis of antibody response to pathogen infection and/or vaccination. However, development of mapping strategies has lagged behind mAb discovery. We have developed a site-directed mutagenesis approach that can be used in conjunction with bio-layer interferometry (BLI) biosensors to map mAb epitopes. By generating a panel of single point mutants in the recombinant hemagglutinin (HA) and neuraminidase (NA) proteins of influenza A viruses, we have characterized the epitopes of hundreds of mAbs targeting the H1 and H3 subtypes of HA and the N9 subtype of NA.
摘要
抗原表位的鉴定对于治疗性抗体、诊断工具和疫苗的开发非常有用。表位作图还为基于序列的病原体感染和/或疫苗接种后抗体反应的库分析提供了功能信息。然而，作图策略的发展落后于单克隆抗体的发现。我们开发了一种定点突变方法，可与生物层干涉（BLI）生物传感器结合使用，以绘制单克隆抗体的表位。通过在甲型流感病毒的重组血凝素（HA）和神经氨酸酶（NA）蛋白中生成一组单点突变体，我们鉴定了针对 HA 的 H1 和 H3 亚型以及 NA 的 N9 亚型的数百种 mAb 的表位。
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