title: Novel human recombinant antibodies against Mycobacterium tuberculosis antigen 85B.
abstract: BACKGROUND
  Tuberculosis is the leading cause of death due to bacterial infections worldwide, mainly caused by Mycobacterium tuberculosis. The antigen 85 complex comprises a set of major secreted proteins of M. tuberculosis, which are potential biomarkers for diagnostic.
  
  
  RESULTS
  In this work, the first human single chain fragment variable (scFv) antibodies specific for the tuberculosis biomarker 85 B were selected by phage display from naïve antibody gene libraries (HAL7/8). Produced as scFv-Fc in mammalian cells, these antibodies were further characterized and analysed for specificity and applicability in different tuberculosis antigen detection assays. Sandwich detection of recombinant 85 B was successful in enzyme linked immunosorbent assay (ELISA), lateral flow immunoassay and immunoblot. Whereas detection of M. tuberculosis cell extracts and culture filtrates was only possible in direct ELISA and immunoblot assays. It was found that the conformation of 85 B, depending on sample treatment, influenced antigen detection.
  
  
  CONCLUSIONS
  Recombinant antibodies, selected by phage display, may be applicable for 85 B detection in various assays. These antibodies are candidates for the development of future point of care tuberculosis diagnostic kits. Using 85 B as a biomarker, the antigen conformation influenced by sample treatment is important.
authors.forename: Manon
authors.initials: M
authors.lastname: Fuchs
authors.forename: Susanne
authors.initials: S
authors.lastname: Kämpfer
authors.forename: Saskia
authors.initials: S
authors.lastname: Helmsing
authors.forename: Ralf
authors.initials: R
authors.lastname: Spallek
authors.forename: Wulf
authors.initials: W
authors.lastname: Oehlmann
authors.forename: Wiebke
authors.initials: W
authors.lastname: Prilop
authors.forename: Ronald
authors.initials: R
authors.lastname: Frank
authors.forename: Stefan
authors.initials: S
authors.lastname: Dübel
authors.forename: Mahavir
authors.initials: M
authors.lastname: Singh
authors.affiliation: Technische Universität Braunschweig, Institut für Biochemie und Biotechnologie, Spielmannstr,7, 38106 Braunschweig, Germany. m.hust@tu-bs.de.
authors.forename: Michael
authors.initials: M
authors.lastname: Hust
mesh_terms: D000217:Acyltransferases / Q000032:analysis / Q000737:chemistry / Q000276:immunology*; D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000032:analysis / Q000737:chemistry / Q000276:immunology*; D001426:Bacterial Proteins / Q000032:analysis / Q000737:chemistry / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D015723:Gene Library; D006801:Humans; D009169:Mycobacterium tuberculosis / Q000378:metabolism*; D019151:Peptide Library; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D000939:Epitopes; D019151:Peptide Library; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D000217:Acyltransferases; C121433:antigen 85B, Mycobacterium tuberculosis