title: Identifying human milk glycans that inhibit norovirus binding using surface plasmon resonance.
abstract: Human milk glycans inhibit binding between norovirus and its host glycan receptor; such competitive inhibition by human milk glycans is associated with a reduced risk of infection. The relationship between the presence of specific structural motifs in the human milk glycan and its ability to inhibit binding by specific norovirus strains requires facile, accurate and miniaturized-binding assays. Toward this end, a high-throughput biosensor platform was developed based on surface plasmon resonance imaging (SPRi) of glycan microarrays. The SPRi was validated, and its utility was tested, by measuring binding specificities between defined human milk glycan epitopes and the capsids of two common norovirus strains, VA387 and Norwalk. Human milk oligosaccharide (HMOS)-based neoglycoconjugates, including chemically derived neoglycoproteins and oligosaccharide-glycine derivatives, were used to represent polyvalent glycoconjugates and monovalent oligosaccharides, respectively, in human milk. SPRi binding results established that the glycan motifs that bind norovirus capsids depend upon strain; VA387 capsid interacts with two neoglycoproteins, whereas Norwalk capsid binds to a different set of HMOS motifs in the form of both polyvalent neoglycoproteins and monovalent oligosaccharides. SPRi competitive binding assays further demonstrated that specific norovirus-binding glycans are able to inhibit norovirus capsid binding to their host receptors. A polyvalent neoglycoconjugate with clustered carbohydrate moieties is required for the inhibition of VA387 capsid binding to host receptor glycans, whereas both monovalent oligosaccharides and polyvalent neoglycoconjugates are able to inhibit Norwalk capsid binding to its host receptor. Binding of HMOS and HMOS-based neoglycoconjugates to norovirus capsids depends upon the specific strain characteristics, implying that HMOS and their polyvalent derivatives are potential anti-adhesive agents for norovirus prophylaxis.
authors.affiliation: Department of Bioengineering, University of Washington, 3720 15th Avenue NE, Seattle, WA 98195, USA.
authors.forename: Jing
authors.initials: J
authors.lastname: Shang
authors.forename: Vladimir E
authors.initials: VE
authors.lastname: Piskarev
authors.forename: Ming
authors.initials: M
authors.lastname: Xia
authors.forename: Pengwei
authors.initials: P
authors.lastname: Huang
authors.forename: Xi
authors.initials: X
authors.lastname: Jiang
authors.forename: Leonid M
authors.initials: LM
authors.lastname: Likhosherstov
authors.forename: Olga S
authors.initials: OS
authors.lastname: Novikova
authors.forename: David S
authors.initials: DS
authors.lastname: Newburg
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Ratner
mesh_terms: D000818:Animals; D001665:Binding Sites / Q000187:drug effects; D002213:Capsid / Q000378:metabolism; D006801:Humans; D008826:Microbial Sensitivity Tests; D008892:Milk / Q000737:chemistry*; D029322:Norovirus / Q000187:drug effects* / Q000378:metabolism; D011134:Polysaccharides / Q000032:analysis* / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D020349:Surface Plasmon Resonance
chemical_list: D011134:Polysaccharides
keywords: anti-adhesives; host–pathogen interactions; human milk glycans; norovirus; surface plasmon resonance
references.citation: J Virol. 2007 Jun;81(11):5949-57
references.pmid: 17392366
references.citation: Expert Rev Mol Med. 2007 Jul 11;9(19):1-22
references.pmid: 17623487
references.citation: Gastroenterology. 2002 Jun;122(7):1967-77
references.pmid: 12055602
references.citation: Arch Biochem Biophys. 1974 Jul;163(1):426-8
references.pmid: 4859528
references.citation: Annu Rev Nutr. 2005;25:37-58
references.pmid: 16011458
references.citation: J Infect Dis. 2008 Sep 15;198(6):940-1; author reply 942-3
references.pmid: 18721066
references.citation: Antimicrob Agents Chemother. 2007 Jan;51(1):324-31
references.pmid: 17060523
references.citation: Chemistry. 2011 Jun 27;17(27):7442-53
references.pmid: 21567493
references.citation: J Virol. 2005 Jun;79(11):6714-22
references.pmid: 15890909
references.citation: Glycobiology. 2013 Mar;23(3):276-85
references.pmid: 23118206
references.citation: Chang Gung Med J. 2005 Aug;28(8):530-42
references.pmid: 16265843
references.citation: J Biol Chem. 1992 Jun 15;267(17):11917-22
references.pmid: 1601861
references.citation: J Virol. 2008 Nov;82(21):10756-67
references.pmid: 18701592
references.citation: J Biochem. 2007 Oct;142(4):459-69
references.pmid: 17652328
references.citation: Biochem J. 2006 Feb 1;393(Pt 3):627-34
references.pmid: 16266293
references.citation: Virology. 2008 Dec 5;382(1):115-23
references.pmid: 18926552
references.citation: Biochemistry. 2001 Mar 13;40(10):3009-15
references.pmid: 11258914
references.citation: Biochem J. 2004 Dec 1;384(Pt 2):307-16
references.pmid: 15361072
references.citation: Glycobiology. 2009 Nov;19(11):1176-84
references.pmid: 19625485
references.citation: Chemistry. 2011 Jun 27;17(27):7438-41
references.pmid: 21469230
references.citation: Org Lett. 2004 Oct 28;6(22):4001-4
references.pmid: 15496084
references.citation: Anal Chem. 2006 Jul 15;78(14):4786-93
references.pmid: 16841896
references.citation: PLoS Pathog. 2010 Aug 26;6(8):e1000983
references.pmid: 20865168
references.citation: J Nutr. 2005 May;135(5):1304-7
references.pmid: 15867329
references.citation: Langmuir. 2012 Jul 31;28(30):11265-73
references.pmid: 22708701
references.citation: J Infect Dis. 2003 Jul 1;188(1):19-31
references.pmid: 12825167
references.citation: ChemMedChem. 2006 Dec;1(12):1361-6
references.pmid: 17042040
references.citation: J Virol. 2003 Dec;77(23):12562-71
references.pmid: 14610179
references.citation: Bioconjug Chem. 2011 Jan 19;22(1):50-7
references.pmid: 21142056
references.citation: J Infect Dis. 2004 Nov 15;190(10):1850-9
references.pmid: 15499543
references.citation: J Infect Dis. 2005 Sep 15;192(6):1071-7
references.pmid: 16107962
references.citation: J Virol. 2008 Jun;82(11):5340-7
references.pmid: 18385236
references.citation: Langmuir. 2012 Feb 14;28(6):3338-44
references.pmid: 22220731
references.citation: Anal Chem. 2004 Dec 15;76(24):7257-62
references.pmid: 15595867
references.citation: Methods. 2000 Sep;22(1):77-91
references.pmid: 11020321
references.citation: Langmuir. 2007 Jan 16;23(2):582-8
references.pmid: 17209609
references.citation: Glycoconj J. 2009 Nov;26(8):1085-96
references.pmid: 19238539
references.citation: Biochemistry. 1974 Jul 16;13(15):3184-92
references.pmid: 4210157