A highly specific monoclonal antibody against monkeypox virus detects the heparin binding domain of A27.

Laura J Hughes, Jason Goldstein, Jan Pohl, Jay W Hooper, R Lee Pitts, Michael B Townsend, Dennis Bagarozzi, Inger K Damon, Kevin L Karem

Virology 2014 Sep

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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. Published by Elsevier Inc.

Citation

Laura J Hughes, Jason Goldstein, Jan Pohl, Jay W Hooper, R Lee Pitts, Michael B Townsend, Dennis Bagarozzi, Inger K Damon, Kevin L Karem. A highly specific monoclonal antibody against monkeypox virus detects the heparin binding domain of A27. Virology. 2014 Sep;464-465:264-273