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Novel human recombinant antibodies against Mycobacterium tuberculosis antigen 85B.

BMC biotechnology (2014-07-19)
Manon Fuchs, Susanne Kämpfer, Saskia Helmsing, Ralf Spallek, Wulf Oehlmann, Wiebke Prilop, Ronald Frank, Stefan Dübel, Mahavir Singh, Michael Hust
ABSTRACT

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. 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. 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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Human IgG (Fc specific)−Peroxidase antibody produced in goat, affinity isolated antibody
Sigma-Aldrich
Anti-Mouse IgG (Fc specific)–Peroxidase antibody produced in goat, affinity isolated antibody

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