Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 411532
Title A Broad Set of Different Llama Antibodies Specific for a 16kDa Heat Shock Protein of Mycobacterium tuberculosis
Author(s) Trilling, A.K.; Ronde, H. de; Noteboom, L.; Houwelingen, A.M.M.L. van; Roelse, M.; Srivastava, S.K.; Haasnoot, W.; Jongsma, M.A.; Kolk, A.; Zuilhof, H.; Beekwilder, J.
Source PLoS One 6 (2011)10. - ISSN 1932-6203 - 10 p.
Department(s) PRI BIOS Applied Metabolic Systems
Laboratory for Organic Chemistry
Publication type Refereed Article in a scientific journal
Publication year 2011
Keyword(s) b-cell - monoclonal-antibodies - rapid detection - in-vitro - antigen - single - fragments - resistance - epitopes - binding
Abstract Background Recombinant antibodies are powerful tools in engineering of novel diagnostics. Due to the small size and stable nature of llama antibody domains selected antibodies can serve as a detection reagent in multiplexed and sensitive assays for M. tuberculosis. Methodology/Principal Findings Antibodies for Mycobacterium tuberculosis (M. tb) recognition were raised in Alpaca, and, by phage display, recombinant variable domains of heavy-chain antibodies (VHH) binding to M. tuberculosis antigens were isolated. Two phage display selection strategies were followed: one direct selection using semi-purified protein antigen, and a depletion strategy with lysates, aiming to avoid cross-reaction to other mycobacteria. Both panning methods selected a set of binders with widely differing complementarity determining regions. Selected recombinant VHHs were produced in E. coli and shown to bind immobilized lysate in direct Enzymelinked Immunosorbent Assay (ELISA) tests and soluble antigen by surface plasmon resonance (SPR) analysis. All tested VHHs were specific for tuberculosis-causing mycobacteria (M. tuberculosis, M. bovis) and exclusively recognized an immunodominant 16 kDa heat shock protein (hsp). The highest affinity VHH had a dissociation constant (KD) of 4×10-10 M. Conclusions/Significance A broad set of different llama antibodies specific for 16 kDa heat shock protein of M. tuberculosis is available. This protein is highly stable and abundant in M. tuberculosis. The VHH that detect this protein are applied in a robust SPR sensor for identification of tuberculosis-causing mycobacteria.
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