The first SPR sensor for detection of bacteria was reported in 1998 with high detection limit as much as 107 cfu/mL. Since then, a lot of effort has been made to lower detection limit and increase sensitivity of detection mainly by using of different assay formats, immobilization strategies, suitable antibodies, minimizing non-specific adsorption and improving the quality of SPR devices. The aim of this short communication is to introduce the potential of anti-OmpW in sensitive detection of V. cholerae by developing an immunosensor based on SPR and compare the sensitivity of this method with former report for detection of V. cholerae published in 2006. Recombinant OmpW antigen (a bacterial outer-membrane protein) of V. cholerae was expressed and purified and raising of polyclonal rabbit anti-OmpW was done. Protein G was covalently immobilized on 11-MUA SAM via amine coupling and bioaffinity-based oriented immobilization of an antibody against recombinant outer membrane protein (anti-OmpW) of Vibrio cholerae was done on protein G layer. The results showed high affinity interaction between OmpW and anti-OmpW (KD=2.4×10-9 M) and the detection limit of fabricated immunosensor was 43 cells/mL. The apparent reasons for achieving this low LOD are discussed.
کلید واژگان :Surface Plasmon Resonance; oriented immobilization; immunosensor; Vibrio cholerae; detection
ارزش ریالی : 500000 ریال
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