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...

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Bibliographic Details
Published in:Biosensors & bioelectronics 2008-10, Vol.24 (2), p.284-289
Main Authors: Wąsowicz, Michał, Viswanathan, Subramanian, Dvornyk, Anzhela, Grzelak, Krystyna, Kłudkiewicz, Barbara, Radecka, Hanna
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biosensing Techniques - instrumentation
Biosensors
Biotechnology
Culture Media - metabolism
Electrochemistry - instrumentation
Equipment Design
Equipment Failure Analysis
Fundamental and applied biological sciences. Psychology
Gold - chemistry
Gold nanorods
Histidine
Histidine - analysis
Immunoassay - instrumentation
Immunoblot
Immunoblotting - instrumentation
Immunosensor
Impedance
Methods. Procedures. Technologies
Nanostructures - chemistry
Nanostructures - ultrastructure
Pichia
Proteins - analysis
Reproducibility of Results
Sensitivity and Specificity
Staining and Labeling - methods
Various methods and equipments
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Summary: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-His6) 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 10pg/ml and 1ng/ml with limit of detection 5pg/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.
ISSN:0956-5663
1873-4235
DOI:10.1016/j.bios.2008.04.002