1. Surface Plasmon Resonance Investigations of Bioselective Element Based on the Recombinant Protein A for Immunoglobulin Detection
- Author
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A. P. Soldatkin, A. E. Rachkov, R. Khristosenko, B. Dons’koi, Iu. V. Ushenin, O. B. Gorbatiuk, A. O. Bakhmachuk, and V. Peshkova
- Subjects
0301 basic medicine ,Materials science ,food.ingredient ,Calibration curve ,Analytical chemistry ,01 natural sciences ,Gelatin ,law.invention ,03 medical and health sciences ,food ,Materials Science(all) ,law ,Surface plasmon resonance ,Immunoglobulin ,General Materials Science ,Recombinant Staphylococcal protein A ,Chromatography ,Nano Express ,010401 analytical chemistry ,Sorption ,Condensed Matter Physics ,0104 chemical sciences ,Affinity biosensor ,030104 developmental biology ,Protein immobilization ,Recombinant DNA ,Selectivity ,Biosensor ,Cysteine - Abstract
The developed surface plasmon resonance (SPR) biosensor based on the recombinant Staphylococcal protein A with an additional cysteine residue (SPA-Cys) used as a biorecognition component showed a good selectivity and sensitivity for the immunoglobulin detection. The developed biosensor with SPA-Cys-based bioselective element can also be used as a first step of immunosensor creation. The successful immobilization of SPA-Cys on the nanolayer gold sensor surface of the SPR spectrometer was performed. The efficiency of blocking nonspecific sorption sites on the sensor surface with milk proteins, gelatin, BSA, and HSA was studied, and a rather high efficiency of using gelatin was confirmed. The SPR biosensor selectively interacted with IgG and did not interact with the control proteins. The linear dependence of the sensor response on the IgG concentration in the range from 2 to 10 μg/ml was shown. Using the calibration curve, the IgG concentration was measured in the model samples. The determined concentrations are in good agreement (r 2 = 0.97) with the given concentration of IgG.
- Published
- 2016