Comparison of electrochemical immunosensors based on gold nano materials and immunoblot techniques for detection of histidine-tagged proteins in culture medium.
In this work, the direct electrochemical determination of poly-histidine tagged proteins using immunosensor based on anti-His (C-term) antibody immobilized on gold electrodes modified with 1,6-hexanedithiol, gold colloid particles or gold nanorods is described. The recombinant histidine-tagged silk proteinase inhibitor protein (rSPI2-His(6)) expressed in Pichia system selected as antigen for this immonosensor. An electrochemical impedance spectroscopy was used as label free detection technique for immune conjugation. The gold nanorods modified electrode layer showed better analytical response than gold nano particles. The linear calibration range was observed between 10 pg/ml and 1 ng/ml with limit of detection 5 pg/ml (S/N=3). Up to four successive assay cycles with retentive sensitivity were achieved for the immunosensors regenerated with 0.2M glycine-HCl buffer, pH 2.8. The performance of this immnosensor were compared with immuoblotting techniques.
Wasowicz
Michał
M
Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Tuwima 10, 10-747 Olsztyn, Poland.
Viswanathan
Subramanian
S
Dvornyk
Anzhela
A
Grzelak
Krystyna
K
Kłudkiewicz
Barbara
B
Radecka
Hanna
H
D015374:Biosensing Techniques / Q000295:instrumentation*; D003470:Culture Media / Q000378:metabolism; D004563:Electrochemistry / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006046:Gold / Q000737:chemistry*; D006639:Histidine / Q000032:analysis*; D007118:Immunoassay / Q000295:instrumentation*; D015151:Immunoblotting / Q000295:instrumentation*; D049329:Nanostructures / Q000737:chemistry* / Q000648:ultrastructure; D011506:Proteins / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods
D003470:Culture Media; D011506:Proteins; D006639:Histidine; D006046:Gold