Adhikari Jagat, Zhao Haiyan, Fernandez Estefania, Huang Yining, Diamond Michael S, Fremont Daved H, Gross Michael L
Department of Chemistry, Washington University in St. Louis, Saint Louis, Missouri, USA.
Department of Pathology and Immunology, Washington University School of Medicine, Saint Louis, Missouri, USA.
J Mass Spectrom. 2021 Jan;56(1). doi: 10.1002/jms.4685. Epub 2020 Nov 16.
Zika Virus (ZIKV) has become a global public health concern because it causes fetal microcephaly and other neurological complications in humans. Currently, there are no approved treatments or vaccines for ZIKV infection. We describe here the detailed epitopes for six monoclonal antibodies (mAbs) that bind to domain III of the envelope protein of ZIKV, some of which have therapeutic potential. We show that by using hydrogen-deuterium exchange mass spectrometry (HDX-MS), we can identify three spatially distinct epitopes for the six mAbs investigated. The HDX-MS approach identified epitopes for three mAbs that agreed well with recently reported X-ray crystallography data. The HDX-MS determined epitopes for the other three anti-ZIKV mAbs for which there were no crystal structures, and the epitopes were confirmed by structure-guided mutagenesis and biolayer interferometry (BLI) competition binding assay. Our results have implications for the design of vaccine and antibody therapeutics against ZIKV and demonstrate the use of HDX-MS as a rapid and valid approach for epitope mapping.
寨卡病毒（ZIKV）已成为全球公共卫生关注的焦点，因为它会导致人类胎儿小头畸形和其他神经并发症。目前，尚无获批用于治疗寨卡病毒感染的药物或疫苗。我们在此描述了六种单克隆抗体（mAb）与寨卡病毒包膜蛋白结构域III结合的详细表位，其中一些具有治疗潜力。我们表明，通过使用氢-氘交换质谱法（HDX-MS），我们可以为所研究的六种单克隆抗体确定三个空间上不同的表位。HDX-MS方法确定的三种单克隆抗体的表位与最近报道的X射线晶体学数据高度吻合。HDX-MS确定了另外三种没有晶体结构的抗寨卡病毒单克隆抗体的表位，这些表位通过结构导向诱变和生物层干涉术（BLI）竞争结合试验得到了证实。我们的结果对寨卡病毒疫苗和抗体疗法的设计具有启示意义，并证明了HDX-MS作为一种快速有效的表位作图方法的应用。
