Novel human recombinant antibodies against Mycobacterium tuberculosis antigen 85B

BMC Biotechnology

Research Article

Ronald Frank¹  Mahavir Singh²  Wulf Oehlmann²  Wiebke Prilop²  Ralf Spallek²  Susanne Kämpfer²  Stefan Dübel³  Manon Fuchs³  Saskia Helmsing³  Michael Hust³ 

[1] Helmholtz Zentrum für Infektionsforschung, Abteilung Chemische Biologie, Inhoffenstr. 7, 38124, Braunschweig, Germany;Leibniz-Institut für Molekulare Pharmakologie, Robert-Roessle-Str. 10, 13125, Berlin, Germany;Lionex GmbH, Salzdahlumer Str. 196, 38126, Braunschweig, Germany;Technische Universität Braunschweig, Institut für Biochemie und Biotechnologie, Spielmannstr.7, 38106, Braunschweig, Germany;

关键词: Culture Filtrate;    Phage Display;    Titration ELISA;    Lateral Flow Immuno Assay;    Tape Station;   

DOI  :  10.1186/1472-6750-14-68

 received in 2014-04-22, accepted in 2014-07-01,  发布年份 2014

来源: Springer

【 摘 要 】

BackgroundTuberculosis 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.ResultsIn 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.ConclusionsRecombinant 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.

【 授权许可】

Unknown   
© Fuchs et al.; licensee BioMed Central Ltd. 2014. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.