The determination of total protein in plant samples is a difficult task, as classical nitrogen-based methods are not selective for the nature of nitrogen, and the results of biochemical methods are influenced by both associated compounds and the complex composition of the protein matrix. Using electrophoretic separation of three commercial sunflower protein samples, it was determined that the studied proteins are a mixture of salt-soluble globulins and water-soluble albumins of different molecular weights. The total protein content of the studied samples was determined using five spectrophotometric methods: direct spectrophotometry, bicinchoninic acid assay, and Benedict’s, Bradford’s, and Lowry’s methods. After comparing the results obtained, it was concluded that, for the determination of protein in these plant materials, the use of the Dumas nitrogen-based method in tandem with Lowry’s spectrophotometric method is the most suitable.
The recent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has posed a great challenge for the development of ultra-fast methods for virus identification based on sensor principles. We created a structure modeling surface and size of the SARS-CoV-2 virus and used it in comparison with the standard antigen SARS-CoV-2—the receptor-binding domain (RBD) of the S-protein of the envelope of the SARS-CoV-2 virus from the Wuhan strain—for the development of detection of coronaviruses using a DNA-modified, surface-enhanced Raman scattering (SERS)-based aptasensor in sandwich mode: a primary aptamer attached to the plasmonic surface—RBD-covered Ag nanoparticle—the Cy3-labeled secondary aptamer. Fabricated novel hybrid plasmonic structures based on “Ag mirror-SiO2-nanostructured Ag” demonstrate sensitivity for the detection of investigated analytes due to the combination of localized surface plasmons in nanostructured silver surface and the gap surface plasmons in a thin dielectric layer of SiO2 between silver layers. A specific SERS signal has been obtained from SERS-active compounds with RBD-specific DNA aptamers that selectively bind to the S protein of synthetic virion (dissociation constants of DNA-aptamer complexes with protein in the range of 10 nM). The purpose of the study is to systematically analyze the combination of components in an aptamer-based sandwich system. A developed virus size simulating silver particles adsorbed on an aptamer-coated sensor provided a signal different from free RBD. The data obtained are consistent with the theory of signal amplification depending on the distance of the active compound from the amplifying surface and the nature of such a compound. The ability to detect the target virus due to specific interaction with such DNA is quantitatively controlled by the degree of the quenching SERS signal from the labeled compound. Developed indicator sandwich-type systems demonstrate high stability. Such a platform does not require special permissions to work with viruses. Therefore, our approach creates the promising basis for fostering the practical application of ultra-fast, amplification-free methods for detecting coronaviruses based on SARS-CoV-2.
15 July 2018 in Journal of Pharmaceutical and Biomedical Analysis
A review of publications, mainly for the last 15 years, characterizing the advantages, limitations, and prospects for the development of modern methods and approaches to the determination of...