Abstract
The correlation between precise interactions of influenza A virus hemagglutinins with host cell surface glycans having terminal sialic acids and host range specificity has provoked the development of a high-throughput viral-receptor specificity assay. Here, we describe the use of the virus itself as a specific antibody coupled to enzymes (virus with neuraminidase spikes) for determining its binding specificity to glycans, a strategy that reduces not only the cost but also the tedious steps of adding primary and secondary antibodies and washing between each step. All of the steps, including coating the glycopolymers onto microtiter plates, virus binding, and visual and quantitative detection of fluorescence products that correlate well with the amount of glycan-bound viruses, can be done within 3 h. This simple, rapid, sensitive, and reliable strategy is an ideal method for detection of high-throughput influenza virus receptor-binding preference not only for studies on viral evolution and transmission but also for viral surveillance in pandemic preparedness, leading to efficient prevention and control of the disease.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Sriwilaijaroen N, Suzuki Y (2012) Molecular basis of the structure and function of H1 hemagglutinin of influenza virus. Proc Jpn Acad Ser B Phys Biol Sci 88:226–249
Stevens J, Blixt O, Paulson JC, Wilson IA (2006) Glycan microarray technologies: tools to survey host specificity of influenza viruses. Nat Rev Microbiol 4:857–864
Xu X, Smith CB, Mungall BA, Lindstrom SE, Hall HE, Subbarao K, Cox NJ, Klimov A (2002) Intercontinental circulation of human influenza A(H1N2) reassortant viruses during the 2001−2002 influenza season. J Infect Dis 186:1490–1493
Kilbourne ED (2006) Influenza pandemics of the 20th century. Emerg Infect Dis 12:9–14
Neumann G, Noda T, Kawaoka Y (2009) Emergence and pandemic potential of swine-origin H1N1 influenza virus. Nature 459:931–939
Sriwilaijaroen N, Kondo S, Yagi H, Hiramatsu H, Nakakita S, Yamada A, Ito H, Hirabayashi J, Narimatsu H, Kato K, Suzuki Y (2012) Bovine milk whey for preparation of natural N-glycans: structural and quantitative analysis. Open Glycoscience 5:41–50
Suzuki Y, Asai A, Suzuki T, Hidari K, Murata T, Usui T, Takeda S, Yamada K, Noguchi T (2009) Method for determination of recognition specificity of virus for receptor sugar chain. US Patent 0181362 A1
Blixt O, Head S, Mondala T, Scanlan C, Huflejt ME, Alvarez R, Bryan MC, Fazio F, Calarese D, Stevens J, Razi N, Stevens DJ, Skehel JJ, van Die I, Burton DR, Wilson IA, Cummings R, Bovin N, Wong CH, Paulson JC (2004) Printed covalent glycan array for ligand profiling of diverse glycan binding proteins. Proc Natl Acad Sci U S A 101:17033–17038
Yamada S, Suzuki Y, Suzuki T, Le MQ, Nidom CA, Sakai-Tagawa Y, Muramoto Y, Ito M, Kiso M, Horimoto T, Shinya K, Sawada T, Usui T, Murata T, Lin Y, Hay A, Haire LF, Stevens DJ, Russell RJ, Gamblin SJ, Skehel JJ, Kawaoka Y (2006) Haemagglutinin mutations responsible for the binding of H5N1 influenza A viruses to human-type receptors. Nature 444:378–382
Cointe D, Leroy Y, Chirat F (1998) Determination of the sialylation level and of the ratio α-(2 → 3)/α-(2 → 6) sialyl linkages of N-glycans by methylation and GC/MS analysis. Carbohydr Res 311:51–59
Gambaryan AS, Matrosovich MN (1992) A solid-phase enzyme-linked assay for influenza virus receptor-binding activity. J Virol Methods 39:111–123
Kobasa D, Takada A, Shinya K, Hatta M, Halfmann P, Theriault S, Suzuki H, Nishimura H, Mitamura K, Sugaya N, Usui T, Murata T, Maeda Y, Watanabe S, Suresh M, Suzuki T, Suzuki Y, Feldmann H, Kawaoka Y (2004) Enhanced virulence of influenza A viruses with the haemagglutinin of the 1918 pandemic virus. Nature 431:703–707
Totani K, Kubota T, Kuroda T, Murata T, Hidari KI, Suzuki T, Suzuki Y, Kobayashi K, Ashida H, Yamamoto K, Usui T (2003) Chemoenzymatic synthesis and application of glycopolymers containing multivalent sialyloligosaccharides with a poly(l-glutamic acid) backbone for inhibition of infection by influenza viruses. Glycobiology 13:315–326
Suzuki Y, Matsunaga M, Matsumoto M (1985) N-Acetylneuraminyllactosylceramide, GM3-NeuAc, a new influenza A virus receptor which mediates the adsorption-fusion process of viral infection. Binding specificity of influenza virus A/Aichi/2/68 (H3N2) to membrane-associated GM3 with different molecular species of sialic acid. J Biol Chem 260:1362–1365
Suzuki Y, Nagao Y, Kato H, Matsumoto M, Nerome K, Nakajima K, Nobusawa E (1986) Human influenza A virus hemagglutinin distinguishes sialyloligosaccharides in membrane-associated gangliosides as its receptor which mediates the adsorption and fusion processes of virus infection. Specificity for oligosaccharides and sialic acids and the sequence to which sialic acid is attached. J Biol Chem 261:17057–17061
Suzuki Y (1994) Gangliosides as influenza virus receptors. Variation of influenza viruses and their recognition of the receptor sialo-sugar chains. Prog Lipid Res 33:429–457
Suzuki Y, Nakao T, Ito T, Watanabe N, Toda Y, Xu G, Suzuki T, Kobayashi T, Kimura Y, Yamada A et al (1992) Structural determination of gangliosides that bind to influenza A, B, and C viruses by an improved binding assay: strain-specific receptor epitopes in sialo-sugar chains. Virology 189:121–131
Suzuki Y, Matsunaga M, Nagao Y, Taki T, Hirabayashi Y, Matsumoto M (1985) Ganglioside GM1b as an influenza virus receptor. Vaccine 3:201–203
Yamashita S, Yoshida H, Uchiyama N, Nakakita Y, Nakakita S, Tonozuka T, Oguma K, Nishikawa A, Kamitori S (2012) Carbohydrate recognition mechanism of HA70 from Clostridium botulinum deduced from X-ray structures in complexes with sialylated oligosaccharides. FEBS Lett 586:2404–2410
Suzuki T, Horiike G, Yamazaki Y, Kawabe K, Masuda H, Miyamoto D, Matsuda M, Nishimura SI, Yamagata T, Ito T, Kida H, Kawaoka Y, Suzuki Y (1997) Swine influenza virus strains recognize sialylsugar chains containing the molecular species of sialic acid predominantly present in the swine tracheal epithelium. FEBS Lett 404:192–196
Tsuchida A, Kobayashi K, Matsubara N, Muramatsu T, Suzuki T, Suzuki Y (1998) Simple synthesis of sialyllactose-carrying polystyrene and its binding with influenza virus. Glycoconj J 15:1047–1054
Sriwilaijaroen N, Kondo S, Yagi H, Wilairat P, Hiramatsu H, Ito M, Ito Y, Kato K, Suzuki Y (2009) Analysis of N-glycans in embryonated chicken egg chorioallantoic and amniotic cells responsible for binding and adaptation of human and avian influenza viruses. Glycoconj J 26:433–443
Takemae N, Ruttanapumma R, Parchariyanon S, Yoneyama S, Hayashi T, Hiramatsu H, Sriwilaijaroen N, Uchida Y, Kondo S, Yagi H, Kato K, Suzuki Y, Saito T (2010) Alterations in receptor-binding properties of swine influenza viruses of the H1 subtype after isolation in embryonated chicken eggs. J Gen Virol 91:938–948
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Sriwilaijaroen, N., Suzuki, Y. (2014). A Simple Viral Neuraminidase-Based Detection for High-Throughput Screening of Viral Hemagglutinin–Host Receptor Specificity. In: Hirabayashi, J. (eds) Lectins. Methods in Molecular Biology, vol 1200. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1292-6_10
Download citation
DOI: https://doi.org/10.1007/978-1-4939-1292-6_10
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1291-9
Online ISBN: 978-1-4939-1292-6
eBook Packages: Springer Protocols