title: Characterization of anti-myosin monoclonal antibodies.
abstract: The characterization of monoclonal antibodies (MAbs) with regard to reactivity and specificity is important for the successful application as a molecular probe and/or diagnostic reagent. Furthermore, it is recognized that some monoclonal reagents perform well in some assay systems but not others. In this study, the reactivity profiles of two anti-myosin MAbs (H1 and DH2, raised against human cardiac myosin) were evaluated in enzyme-linked immunosorbent assay (ELISA), slot-blotting, and immunocytochemistry. Both antibodies performed well in slot-blotting against myosin heavy chain preparations from cardiac and skeletal muscle and from non-human sources. In general, MAb H1 demonstrated strong to moderate reactivity in all assay systems, whilst MAb DH2 faired poorly in ELISA. MAb H1 also showed reactivity to synthetic peptides of myosin, one of which possessed a motif (ERRDA, single amino acid code) that was found in other human and nonhuman myosin protein sequences that could explain its cross-reactive profile. Intriguingly, this motif was found on viral and other pathogenic agents associated with myocarditis. Hence, it is speculated that this region could give some credence to the mechanism of molecular mimicry associated with some cardiac diseases. Overall, MAb H1 may serve as a useful probe of myosin structure.
authors.affiliation: Molecular Immunology Research Group, Research Institute in Healthcare Science, School of Applied Sciences, University of Wolverhampton, Wolverhampton, United Kingdom. p.n.nelson@wlv.ac.uk
authors.forename: P N
authors.initials: PN
authors.lastname: Nelson
authors.forename: S J
authors.initials: SJ
authors.lastname: Astley
authors.forename: D
authors.initials: D
authors.lastname: Roden
authors.forename: E
authors.initials: E
authors.lastname: Waldron
authors.forename: K
authors.initials: K
authors.lastname: Baig
authors.forename: A L P
authors.initials: AL
authors.lastname: Caforio
authors.forename: Y
authors.initials: Y
authors.lastname: Koutedakis
authors.forename: S
authors.initials: S
authors.lastname: Perera
authors.forename: C
authors.initials: C
authors.lastname: Spry
mesh_terms: D020816:Amino Acid Motifs / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology*; D000918:Antibody Specificity; D024722:Cardiac Myosins / Q000276:immunology*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007150:Immunohistochemistry; D008969:Molecular Sequence Data
chemical_list: D000911:Antibodies, Monoclonal; D024722:Cardiac Myosins