SPR biosensor for the detection of L. monocytogenes using phage-displayed antibody.
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 x 10(6)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.
Nanduri
Viswaprakash
V
Center For Food Safety and Engineering, Department of Food Science, Purdue University, 745 Agriculture Mall Dr., West Lafayette, IN 47907, USA.
Bhunia
Arun K
AK
Tu
Shu-I
SI
Paoli
George C
GC
Brewster
Jeffrey D
JD
D000911:Antibodies, Monoclonal / Q000276:immunology*; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D015169:Colony Count, Microbial / Q000295:instrumentation / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007118:Immunoassay / Q000295:instrumentation* / Q000379:methods; D008089:Listeria monocytogenes / Q000276:immunology* / Q000302:isolation & purification*; D019151:Peptide Library; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance / Q000379:methods*
D000911:Antibodies, Monoclonal; D019151:Peptide Library