title: A highly specific monoclonal antibody against monkeypox virus detects the heparin binding domain of A27.
abstract: The eradication of smallpox and the cessation of global vaccination led to the increased prevalence of human infections in Central Africa. Serologic and protein-based diagnostic assay for MPXV detection is difficult due to cross-reactive antibodies that do not differentiate between diverse orthopoxvirus (OPXV) species. A previously characterized monoclonal antibody (mAb 69-126-3-7) against MPXV [1] was retested for cross-reactivity with various OPXVs. The 14.5 kDa band protein that reacted with mAb 69-126-3 was identified to be MPXV A29 protein (homolog of vaccinia virus Copenhagen A27). Amino acid sequence analysis of the MPXV A29 with other OPXV homologs identified four amino acid changes. Peptides corresponding to these regions were designed and evaluated for binding to mAb 69-126-3 by ELISA and BioLayer Interferometry (BLI). Further refinement and truncations mapped the specificity of this antibody to a single amino acid difference in a 30-mer peptide compared to other OPXV homologs. This particular residue is proposed to be essential for heparin binding by VACV A27 protein. Despite this substitution, MPXV A29 bound to heparin with similar affinity to that of VACV A27 protein, suggesting flexibility of this motif for heparin binding. Although binding of mAb 69-126-3-7 to MPXV A29 prevented interaction with heparin, it did not have any effect on the infectivity of MPXV. Characterization of 69-126-3-7 mAb antibody allows for the possibility of the generation of a serological based species-specific detection of OPXVs despite high proteomic homology.
authors.affiliation: Centers for Disease Control and Prevention, Atlanta, GA, USA. Electronic address: bkz2@cdc.gov.
authors.forename: Laura J
authors.initials: LJ
authors.lastname: Hughes
authors.affiliation: Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Goldstein
authors.affiliation: Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: Jan
authors.initials: J
authors.lastname: Pohl
authors.affiliation: USAMRIID, Frederick, MD, USA.
authors.forename: Jay W
authors.initials: JW
authors.lastname: Hooper
authors.affiliation: Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: R
authors.initials: R
authors.lastname: Lee Pitts
authors.affiliation: Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: Michael B
authors.initials: MB
authors.lastname: Townsend
authors.affiliation: Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: Dennis
authors.initials: D
authors.lastname: Bagarozzi
authors.affiliation: Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: Inger K
authors.initials: IK
authors.lastname: Damon
authors.affiliation: Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: Kevin L
authors.initials: KL
authors.lastname: Karem
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D003429:Cross Reactions; D006493:Heparin / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D045908:Mpox, Monkeypox / Q000276:immunology / Q000821:virology*; D008993:Monkeypox virus / Q000737:chemistry / Q000276:immunology*; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D014764:Viral Proteins / Q000737:chemistry / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D014764:Viral Proteins; D006493:Heparin
keywords: Antibody; Heparin binding; Monkeypox virus; Orthopoxvirus; Vaccinia virus
references.citation: J Mol Biol. 2005 Jun 24;349(5):1060-71
references.pmid: 15913650
references.citation: Vector Borne Zoonotic Dis. 2012 May;12(5):400-9
references.pmid: 22217169
references.citation: J Clin Virol. 2006 Jul;36(3):194-203
references.pmid: 16731033
references.citation: J Biol Chem. 2014 Mar 7;289(10):6639-6655
references.pmid: 24451374
references.citation: J Virol Methods. 2010 Oct;169(1):223-7
references.pmid: 20643162
references.citation: J Virol. 2008 Dec;82(24):12384-91
references.pmid: 18842719
references.citation: J Virol. 2009 Jul;83(13):6464-76
references.pmid: 19369327
references.citation: J Gen Virol. 2000 Jan;81(Pt 1):47-58
references.pmid: 10640541
references.citation: N Engl J Med. 2004 Jan 22;350(4):342-50
references.pmid: 14736926
references.citation: J Virol. 2010 Aug;84(15):7592-602
references.pmid: 20484506
references.citation: J Virol. 1998 Feb;72(2):1577-85
references.pmid: 9445060
references.citation: J Virol Methods. 2011 Aug;175(2):163-9
references.pmid: 21635922
references.citation: J Virol. 1993 Jun;67(6):3435-40
references.pmid: 8497059
references.citation: Nat Med. 2005 Sep;11(9):1005-11
references.pmid: 16086024
references.citation: Virology. 2012 Nov 25;433(2):506-12
references.pmid: 22999097
references.citation: Curr Opin Chem Biol. 2001 Jun;5(3):302-7
references.pmid: 11479122
references.citation: J Virol. 2003 Sep;77(18):9894-905
references.pmid: 12941899
references.citation: Proc Natl Acad Sci U S A. 2010 Sep 14;107(37):16262-7
references.pmid: 20805472
references.citation: Nucleic Acids Res. 1990 Sep 25;18(18):5347-51
references.pmid: 2216706
references.citation: J Virol. 2013 Jan;87(2):900-11
references.pmid: 23135728
references.citation: PLoS One. 2010 Dec 30;5(12):e15547
references.pmid: 21209900
references.citation: Proc Natl Acad Sci U S A. 1981 Jun;78(6):3824-8
references.pmid: 6167991
references.citation: Virology. 1991 Apr;181(2):742-8
references.pmid: 2014647
references.citation: J Mol Biol. 1982 May 5;157(1):105-32
references.pmid: 7108955
references.citation: Proteomics. 2007 May;7(10):1678-86
references.pmid: 17443847
references.citation: Virology. 2008 Jul 20;377(1):19-29
references.pmid: 18482742
references.citation: Cell Microbiol. 2008 Jan;10(1):149-64
references.pmid: 17708756
references.citation: J Virol. 2011 Aug;85(15):7683-98
references.pmid: 21632764
references.citation: J Virol. 1984 Oct;52(1):290-2
references.pmid: 6207310
references.citation: Virology. 1990 Sep;178(1):81-91
references.pmid: 2389560
references.citation: J Clin Microbiol. 2004 May;42(5):1940-6
references.pmid: 15131152
references.citation: J Virol. 1998 Oct;72(10):8374-9
references.pmid: 9733888
references.citation: J Biol Chem. 2009 Dec 25;284(52):36535-36546
references.pmid: 19858217
references.citation: J Virol. 1999 Nov;73(11):9098-109
references.pmid: 10516016
references.citation: J Virol. 2008 Apr;82(7):3751-68
references.pmid: 18234801
references.citation: J Biol Chem. 2002 Jun 7;277(23):20949-59
references.pmid: 11901147