Lancaster Catherine, Pristatsky Pavlo, Hoang Van M, Casimiro Danilo R, Schwartz Richard M, Rustandi Richard, Ha Sha
Vaccine Bioprocess Research & Development, Merck Research Laboratories, West Point, PA, 19486, United States.
Vaccine Production Program, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Gaithersburg, MD 20878, United States.
J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Oct 1;1032:218-223. doi: 10.1016/j.jchromb.2016.04.025. Epub 2016 Apr 21.
Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that causes severe arthralgia. The envelope of CHIKV is composed of 240 copies of two glycoproteins: E1 and E2. In this work, we have characterized the N-glycosylation patterns of CHIKV virus-like particles (VLPs), containing both E1 and E2 proteins, derived from mammalian and insect cells using hydrophilic interaction liquid chromatography (HILIC) with fluorescence (FL) and mass spectrometry (MS) detection. While HEK293 derived CHIKV VLPs contain oligomannose, hybrid and complex glycans, VLPs derived from SfBasic predominantly contain oligomannose glycans. This strong host dependence of N-glycosylation pattern resembles other alphaviruses such as SINV. The VLPs from HEK293 and SfBasic, with significantly different N-glycosylation profiles, are valuable reagents enabling future in-depth correlation studies between immunogenicity and glycosylation. In addition, the characterization tools presented here allow one to monitor glycosylation during vaccine process development and ensure process consistency.
基孔肯雅病毒（CHIKV）是一种由蚊子传播的甲病毒，可引起严重的关节痛。CHIKV的包膜由两种糖蛋白的240个拷贝组成：E1和E2。在这项工作中，我们利用亲水相互作用液相色谱（HILIC）结合荧光（FL）和质谱（MS）检测，对源自哺乳动物和昆虫细胞的含有E1和E2蛋白的CHIKV病毒样颗粒（VLP）的N-糖基化模式进行了表征。虽然源自HEK293的CHIKV VLP含有高甘露糖型、杂合型和复合型聚糖，但源自SfBasic的VLP主要含有高甘露糖型聚糖。N-糖基化模式的这种强烈宿主依赖性类似于其他甲病毒，如辛德毕斯病毒（SINV）。来自HEK293和SfBasic的VLP具有明显不同的N-糖基化谱，是未来进行免疫原性和糖基化之间深入相关性研究的有价值试剂。此外，本文介绍的表征工具可使人们在疫苗工艺开发过程中监测糖基化情况，并确保工艺的一致性。
