PDF(Pubmed)
Abstract：
Antigen-antibody interactions play a key role in the immune response post vaccination and the mechanism of action of antibody-based biopharmaceuticals. 4CMenB is a multicomponent vaccine against Neisseria meningitidis serogroup B in which factor H binding protein (fHbp) is one of the key antigens. In this study, we use hydrogen/deuterium exchange mass spectrometry (HDX-MS) to identify epitopes in fHbp recognized by polyclonal antibodies (pAb) from two human donors (HDs) vaccinated with 4CMenB. Our HDX-MS data reveal several epitopes recognized by the complex mixture of human pAb. Furthermore, we show that the pAb from the two HDs recognize the same epitope regions. Epitope mapping of total pAb and purified fHbp-specific pAb from the same HD reveals that the two antibody samples recognize the same main epitopes, showing that HDX-MS based epitope mapping can, in this case at least, be performed directly using total IgG pAb samples that have not undergone Ab-selective purification. Two monoclonal antibodies (mAb) were previously produced from B-cell repertoire sequences from one of the HDs and used for epitope mapping of fHbp with HDX-MS. The epitopes identified for the pAb from the same HD in this study, overlap with the epitopes recognized by the two individual mAbs. Overall, HDX-MS epitope mapping appears highly suitable for simultaneous identification of epitopes recognized by pAb from human donors and to thus both guide vaccine development and study basic human immunity to pathogens, including viruses.
摘要：
抗原-抗体相互作用在疫苗接种后的免疫应答和基于抗体的生物药物的作用机制中起关键作用。4CMenB是针对脑膜炎奈瑟菌血清群B的多组分疫苗，其中H因子结合蛋白（fHbp）是关键抗原之一。在这项研究中,我们使用氢/氘交换质谱（HDX-MS）来鉴定由来自接种4CMenB的两个人供体（HDs）的多克隆抗体（pAb）识别的fHbp中的表位。我们的HDX-MS数据揭示了由人pAb的复杂混合物识别的几个表位。此外，我们显示来自两个HDs的pAb识别相同的表位区域。来自同一HD的总pAb和纯化的fHbp特异性pAb的表位定位揭示了两种抗体样品识别相同的主表位,表明基于HDX-MS的表位定位可以，至少在这种情况下,这可以例如直接使用未经历Ab选择性纯化的总IgGpAb样品进行。两种单克隆抗体（mAb）先前从来自HDs之一的B细胞库序列产生，并用于用HDX-MS对fHbp进行表位定位。在这项研究中，来自同一HD的pAb鉴定的表位，与由两个单独的mAb识别的表位重叠。总的来说,HDX-MS表位作图似乎非常适合同时鉴定来自人类供体的pAb识别的表位，从而指导疫苗开发和研究人类对病原体的基本免疫力。包括病毒。
