Morrissey Bethny, Streamer Margaret, Downard Kevin M
School of Molecular & Microbial Biosciences, The University of Sydney, Australia.
J Virol Methods. 2007 Nov;145(2):106-14. doi: 10.1016/j.jviromet.2007.05.015. Epub 2007 Jun 22.
The antigenic characterisation of three H3N2 type A influenza strains by mass spectrometry is described. The approach, developed in this laboratory, employs matrix-assisted laser desorption ionisation (MALDI) mass spectrometry to analyse gel-resolved antigens, post their proteolysis and treatment with monoclonal antibodies. The primary structure and antigenicity of the component antigens of the virus can be determined in a single step. Four antigenic domains of hemagglutinin have been identified and these are localised at residues 109-125, 158-170 and 316-326 of the HA1 subunit and to residues 159-183 of the HA2 subunit. The results demonstrate the applicability of the approach for identifying antigenic determinants across various H3N2 strains with high throughput and at low sample levels. Comparative rates of antibody binding between two of the antigenic peptides have also been reported.
本文描述了通过质谱法对三株H3N2型甲型流感病毒株进行抗原表征的过程。该方法由本实验室开发，采用基质辅助激光解吸电离（MALDI）质谱法，在凝胶分离的抗原经蛋白酶解并用单克隆抗体处理后对其进行分析。病毒组成抗原的一级结构和抗原性可在一步中确定。已鉴定出血凝素的四个抗原结构域，它们位于HA1亚基的第109 - 125、158 - 170和316 - 326位氨基酸残基处，以及HA2亚基的第159 - 183位氨基酸残基处。结果表明该方法适用于高通量、低样品量条件下鉴定各种H3N2病毒株的抗原决定簇。还报道了两种抗原肽之间抗体结合的比较速率。
