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SPR biosensor for the detection of L. monocytogenes using phage-displayed antibody

2007; Elsevier BV; Volume: 23; Issue: 2 Linguagem: Inglês

10.1016/j.bios.2007.04.007

1873-4235

Viswaprakash Nanduri, Arun K. Bhunia, Shu‐I Tu, George C. Paoli, Jeffrey D. Brewster,

Advanced Biosensing Techniques and Applications

Whole cells of Listeria monocytogenes were detected with a compact, surface plasmon resonance (SPR) sensor using a phage-displayed scFv antibody to the virulence factor actin polymerization protein (ActA) for biorecognition. Phage Lm P4:A8, expressing the scFv antibody fused to the pIII surface protein was immobilized to the sensor surface through physical adsorption. A locally constructed fluidics system was used to deliver solutions to the compact, two-channel SPREETA™ sensor. Specificity of the sensor was tested using common food-borne bacteria and a control phage, M13K07 lacking the scFv fusion on its coat protein. The detection limit for L. monocytogenes whole cells was estimated to be 2 × 106 cfu/ml. The sensor was also used to determine the dissociation constant (Kd) for the interaction of phage-displayed scFv and soluble ActA in solution as 4.5 nM.