title: Differential recognition of the multiple banded antigen isoforms across Ureaplasma parvum and Ureaplasma urealyticum species by monoclonal antibodies.
abstract: Two separate species of Ureaplasma have been identified that infect humans: Ureaplasma parvum and Ureaplasma urealyticum. Most notably, these bacteria lack a cell wall and are the leading infectious organism associated with infection-related induction of preterm birth. Fourteen separate representative prototype bacterial strains, called serovars, are largely differentiated by the sequence of repeating units in the C-terminus of the major surface protein: multiple-banded antigen (MBA). Monoclonal antibodies that recognise single or small groups of serovars have been previously reported, but these reagents remain sequestered in individual research laboratories. Here we characterise a panel of commercially available monoclonal antibodies raised against the MBA and describe the first monoclonal antibody that cross-reacts by immunoblot with all serovars of U. parvum and U. urealyticum species. We also describe a recombinant MBA expressed by Escherichia coli which facilitated further characterisation by immunoblot and demonstrate immunohistochemistry of paraffin-embedded antigens. Immunoblot reactivity was validated against well characterised previously published monoclonal antibodies and individual commercial antibodies were found to recognise all U. parvum strains, only serovars 3 and 14 or only serovars 1 and 6, or all strains belonging to U. parvum and U. urealyticum. MBA mass was highly variable between strains, consistent with variation in the number of C-terminal repeats between strains. Antibody characterisation will enable future investigations to correlate severity of pathogenicity to MBA isoform number or mass, in addition to development of antibody-based diagnostics that will detect infection by all Ureaplasma species or alternately be able to differentiate between U. parvum, U. urealyticum or mixed infections.
authors.affiliation: Cardiff University, School of Medicine, Institute of Molecular and Experimental Medicine, 5th floor University Hospital of Wales, Cardiff CF14 4XN, UK; Sebha University, Faculty of Engineering and Technology, Medical Laboratory Sciences Department, P.O. Box 68, Brak, Libya.
authors.forename: Ali F
authors.initials: AF
authors.lastname: Aboklaish
authors.affiliation: Cardiff University, School of Medicine, Institute of Molecular and Experimental Medicine, 5th floor University Hospital of Wales, Cardiff CF14 4XN, UK; Department of Pathology, Nineveh College of Medicine, University of Mosul, Mosul, Iraq.
authors.forename: Shatha
authors.initials: S
authors.lastname: Ahmed
authors.affiliation: Virostat Inc., P.O. Box 8522, Portland, ME 04104, USA.
authors.forename: Douglas
authors.initials: D
authors.lastname: McAllister
authors.affiliation: Harvard Medical School, Department of Global Health and Social Medicine, Boston, MA 02115, USA; Infectious Disease Research Institute, Seattle, WA 98102, USA.
authors.forename: Gail
authors.initials: G
authors.lastname: Cassell
authors.affiliation: Ann & Robert H. Lurie Children's Hospital of Chicago and Northwestern University Feinberg School of Medicine, Chicago, IL 60611-2605, USA.
authors.forename: Xiaotian T
authors.initials: XT
authors.lastname: Zheng
authors.affiliation: Cardiff University, School of Medicine, Institute of Molecular and Experimental Medicine, 5th floor University Hospital of Wales, Cardiff CF14 4XN, UK. Electronic address: SpillerB@cardiff.ac.uk.
authors.forename: Owen B
authors.initials: OB
authors.lastname: Spiller
mesh_terms: D000907:Antibodies, Bacterial / Q000276:immunology* / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D001426:Bacterial Proteins / Q000235:genetics; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D015151:Immunoblotting; D007158:Immunologic Techniques; D016133:Polymerase Chain Reaction; D020033:Protein Isoforms / Q000276:immunology; D017422:Sequence Analysis, DNA; D065288:Serogroup; D014509:Ureaplasma / Q000276:immunology*; D016869:Ureaplasma Infections / Q000175:diagnosis / Q000276:immunology / Q000382:microbiology; D016990:Ureaplasma urealyticum / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D020033:Protein Isoforms
keywords: Multiple banded antigen; Recombinant protein; Ureaplasma parvum; Ureaplasma urealyticum