A MODEL OF SURFACE PLASMON RESONANCE IMMUNOSENSOR FOR DETERMINING AUTOANTIBODIES TO GLUTATHIONE-S-TRANSFERASE IN SERUM SAMPLES

DOI: https://doi.org/10.29296/24999490-2020-06-06

P.V. Ershov, L.A. Kaluzhskiy, A.S. Ivanov Institute of Biomedical Chemistry, Pogodinskaya str., 10, Bld. 8, Moscow, 119121, Russian Federation E-mail: [email protected]

Introduction. The quantitative determination and isotyping of autoantibodies to a clinically significant protein – glutathione-S-transferase (GST) in blood serum samples is relevant for monitoring and predicting the dynamics of autoimmune processes. The aim of this study. To adapt an optical biosensor based on surface plasmon resonance (SPR) to detect the interactions of GST and its antibodies in several model experiments using blood serum samples. Methods. The SPR method was used to detect antigen-antibody complexes in the direct, signal enhancement and competitive inhibition assays. Results. For the first time it was shown that there is a principle possibility of SPR analysis for the detection of polyclonal antibodies against GST as an antigen in blood serum samples and isotype determination of bound antibodies with the following immunosensor parameters: a) the immobilization level of GST on a CM5 chip was 4–4.5 ng/mm2; b) the ligand binding capacity decrease after each biosensor cycle, when using 10 mM glycine-HCl (pH 1.5) as an regeneration solution, was 0.1–0.3%; c) linearity range was 30–325 nM; d) the serum content in the analyzed samples was 10% (v/v). Conclusion. Thus, the adaptation of the Biacore SPR biosensors (GE Healthcare, USA) to serial analysis of serum samples to monitor the presence and dynamics of autoantibodies to human body proteins (including GST) opens up new ways for the development of diagnostic tools in biomedical and clinical studies.
Keywords: 
surface plasmon resonance (SPR), isotyping, antibodies, experimental model, optical biosensor, glutathione-S-transferase
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