Molecular diagnostics based on surface plasmon angular spectroscopy

• Дата: 2020
• Автори: Kostyukevych, K. V., Khrystosenko, R. V., Zagorodnya, S. D., Kostyukevych, S. O., Koptiukh, A. A., Kryuchyn, A. A., Oleksenko, P. F.
• Формат: Стаття
• Мова: Ukrainian
• Опубліковано: Інститут проблем реєстрації інформації НАН України 2020
• Теми: поверхневий плазмонний резонанс; антитіла; міоглобін; фібриноген; віруси; буферні розчини; плазма та сироватка крові; surface plasmon resonance; antibodies; viruses; myoglobin; fibrinogen; buffer solutions; blood plasma and serum
• Онлайн доступ: http://drsp.ipri.kiev.ua/article/view/218824
• Назва журналу: Data Recording, Storage & Processing

With the purpose to reliably detect specific antibodies against the Epstein–Barr virus (EBV) and cardiovascular system molecular markers (myoglobin and fibrinogen) levels in the buffer solutions, human blood plasma and serum, it has been analyzed some techniques for protein immobilization from immune pair (antibody, antigen) onto the gold surface, as well as for increasing of sensor device («Plasmon» series instrument) sensitivity, stability and productivity, which based on surface plasmon angular spectroscopy with prism excitation.Immobilization of the EBV proteins was performed within the protective-orienting coating — three-dimensional structure of the polysaccharide dextran hydrogel deposited onto the gold film surface on glass substrate, the remaining free sites were blocked with bovine serum albumin molecules (1% BSA) to prevent non-specific interaction, and test the human blood serum from patient with EBV infection under 1:100 delusion in citrate buffer. For oriented attaching of monoclonal antibodies of  mouse against the human myoglobin on the surface of gold protein A Staphylococcus aureus conjugate was used, and carry out investigation in human blood serum without delusion. The procedure of formation of am inorganic buffer layer on the surface of a chip for the site-oriented immobilization of biological receptors with the use of the step-like self-organization of the complex ferrocyanides of transition metals (five-ply film of copper aminopentacyanoferrate Cu3 [Fe(CN)5NH3]2) is developed. This gives possibility to get the concentration and calibrating curves (maximum sensitivity of 0,5 μg/ml) for the model fibrinogen in phosphate buffer, and to determine the concentration of fibrinogen in donors blood plasma under 1:300.A four-channel modification of the «Plasmon» series device with successive inquiry of photodiodes massif and with application of comparison reference channel to minimize the effect of the temperature change induced noise has been constructed for the laboratory studies execution. For the large-scale testing of the population, we propose a construction of the sensor in disk format on the basis of the prism scheme of excitation of SPR in the Kretschmann configuration with optical scanning over angles, where a planar chip with integrated gratings for the input-output of light was used as a sensor unit. Fig.: 14. Refs: 45 titles.