a broad set of different llama antibodies specific for a 16 kda heat shock protein of mycobacterium tuberculosis一系列的不同的骆驼抗体特定16 kda热休克蛋白的结核分枝杆菌.pdfVIP

A Broad Set of Different Llama Antibodies Specific for a 16 kDa Heat Shock Protein of Mycobacterium tuberculosis 1,2 3 1 ` 1 1 Anke K. Trilling , Hans de Ronde , Linda Noteboom , Adele van Houwelingen , Margriet Roelse , 1 5 1 4 2 Saurabh K. Srivastava , Willem Haasnoot , Maarten A. Jongsma , Arend Kolk , Han Zuilhof , Jules Beekwilder1* 1 Plant Research International, Wageningen, The Netherlands, 2 Laboratory of Organic Chemistry, Wageningen University, Wageningen, The Netherlands, 3 Royal Tropical Institute, Amsterdam, The Netherlands, 4 Van ’t Hoff Institute for Molecular Sciences, University of Amsterdam, Amsterdam, The Netherlands, 5 RIKILT-Institute of Food Safety, Wageningen, The Netherlands 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 immobili