Glycoscience & Glycotechnology Research Group, Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science & Technology, Tsukuba, Ibaraki 305-8568, Japan.
Department of Biochemistry, School of Medicine, Keio University, Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.
Virology. 2019 Jan 15;527:132-140. doi: 10.1016/j.virol.2018.11.010. Epub 2018 Nov 29.
Glycan structures on hemagglutinin (HA) of influenza A viruses have been analyzed previously to understand their significance. However, the formerly established methods using mass spectrometry present disadvantages such as procedure complexity, sensitivity, and throughput. Our study has established a novel method for analyzing glycan profiles of HA using lectin microarray techniques. We successfully obtained glycan profiles of HA starting from 1 ml of the 10 TCID samples through simple antigen enrichment using optimized immunoprecipitation. The profiles were reasonably consistent with known glycan structures of HA. Next, we compared glycan profiles of the HAs prepared from chicken embryos, MDCK, Vero, and A549 cells, and demonstrated the host cell-specific HA glycan profiles. Notably, the HA from MDCK cells was α1-3 galactosylated. Our method provides a highly sensitive and simple procedure for glycan profiling of the viral glycoproteins, thereby paving way for direct glycan analyses of human- and animal-derived virions.
先前已经分析了甲型流感病毒血凝素 (HA) 上的聚糖结构，以了解其重要性。然而，以前使用质谱法建立的方法存在程序复杂、灵敏度和通量等缺点。我们的研究建立了一种使用凝集素微阵列技术分析 HA 聚糖图谱的新方法。我们通过使用优化的免疫沉淀进行简单的抗原富集，成功地从 10 TCID 的 1 毫升样本中获得了 HA 的聚糖图谱。这些图谱与已知的 HA 聚糖结构相当一致。接下来，我们比较了从鸡胚、MDCK、Vero 和 A549 细胞中制备的 HA 的聚糖图谱，并证明了宿主细胞特异性的 HA 聚糖图谱。值得注意的是，MDCK 细胞中的 HA 被α1-3 半乳糖化。我们的方法为病毒糖蛋白的聚糖分析提供了一种高度敏感和简单的程序，从而为直接分析人源和动物源病毒粒子的聚糖铺平了道路。
