Study on interactions of human IgG with immobilized anti-IgG or recombinant Staphylococcal protein A using surface plasmon resonance spectrometry

• Published in: Biopolimery i kletka 2016, Vol.32 (1), p.54-60
• Main Authors: Bakhmachuk, A. O., Gorbatiuk, O. B., Palyvoda, O. G., Dons'koi, B. V., Rachkov, A. E., Soldatkin, A. P.
• Format: Article
• Language: English
• Subjects: Immobilization; Immunoglobulin G; Pathogens; Protein A; Scientific imaging; Sensors; Spectrometry; Staphylococcus aureus; Surface plasmon resonance
• ISSN: 0233-7657; 1993-6842
• DOI: 10.7124/bc.00090D

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.