Biopolym. Cell. 2016; 32(1):54-60.
http://dx.doi.org/10.7124/bc.00090D
Molecular and Cell Biotechnologies
Study on interactions of human IgG with immobilized anti-IgG or recombinant Staphylococcal protein A using surface plasmon resonance spectrometry
1, 2Bakhmachuk A. O., 1, 3Gorbatiuk O. B., 1, 2, 3Palyvoda O. G., 4Dons'koi B. V., 1Rachkov A. E., 1Soldatkin A. P.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
  2. Educational and Scientific Center "Institute of Biology",
    Taras Shevchenko National University of Kyiv
    64/13, Volodymyrska Str., Kyiv, Ukraine, 01601
  3. State Institute of Genetic and Regenerative Medicine, NAMS of Ukraine
    67, Vyshhorodska Str., Kyiv, Ukraine, 04114
  4. Institute of Pediatrics, Obstetrics and Gynecology, NAMS of Ukraine
    8, Platona Mayborody St., Kyiv, Ukraine, 04050

Abstract

Aim. Comparison of the IgG-binding activity of recombinant Staphylococcal protein A with introduced C-terminal cysteine residue (SPA-Cys) or goat anti-human IgG antibodies (anti-IgG) after their immobilization on a gold sensor surface of surface plasmon resonance (SPR) spectrometer. Methods. SPA-Cys or anti-IgG were immobilized on a gold sensor surface to form two variants of a bioselective element of the immunosensor. SPR spectrometry was used for the detection of IgG-binding activity of the immobilized proteins. Results.The SPR sensor response to the immobilization of anti-IgG was more than two times higher than that at the immobilization of SPA-Cys. However, there is almost the double advantage for SPA-Cys in the number of immobilized molecules. Moreover, the bioselective element of the immunosensor based on SPA-Cys showed a much better capability of binding Ig than bioselective element based on anti-IgG. Conclusions.The study on the immobilization of SPA-Cys or anti-IgG on the sensor surface of SPR spectrometer, and the interactions of immobilized proteins with human IgG demonstrated obvious advantages of SPA-Cys.
Keywords: antibodies, recombinant Staphylococcal protein A, protein immobilization, immunosensor, surface plasmon resonance
Full text: (PDF, in English)

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