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1.
Biomed Khim ; 69(5): 253-280, 2023 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-37937429

RESUMEN

Traditional antiviral vaccines are currently created by inactivating the virus chemically, most often using formaldehyde or ß-propiolactone. These approaches are not optimal since they negatively affect the safety of the antigenic determinants of the inactivated particles and require additional purification stages. The most promising platforms for creating vaccines are based on pseudoviruses, i.e., viruses that have completely preserved the outer shell (capsid), while losing the ability to reproduce owing to the destruction of the genome. The irradiation of viruses with electron beam is the optimal way to create pseudoviral particles. In this review, with the example of the poliovirus, the main algorithms that can be applied to characterize pseudoviral particles functionally and structurally in the process of creating a vaccine preparation are presented. These algorithms are, namely, the analysis of the degree of genome destruction and coimmunogenicity. The structure of the poliovirus and methods of its inactivation are considered. Methods for assessing residual infectivity and immunogenicity are proposed for the functional characterization of pseudoviruses. Genome integrity analysis approaches, atomic force and electron microscopy, surface plasmon resonance, and bioelectrochemical methods are crucial to structural characterization of the pseudovirus particles.


Asunto(s)
Poliomielitis , Poliovirus , Vacunas , Humanos , Poliovirus/genética , Formaldehído , Propiolactona , Poliomielitis/prevención & control
2.
Dokl Biochem Biophys ; 506(1): 215-219, 2022 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-36303055

RESUMEN

A new approach for modifying electrodes using porous membranes based on anodic aluminum oxide with pore diameters of 0.1 and 0.2 µm and a membrane-like substance didodecyldimethylammonium bromide (DDAB) was proposed to study the electrocatalytic efficiency of the system. This approach makes it possible to increase the catalytic efficiency of the cytochrome P450 3A4-dependent N-demethylation of erythromycin by 132% when using a membrane with pore diameter of 0.1 µm and by 32% when using a membrane with 0.2 µm pore size. Electrode modification using porous membranes shifted the potential of electrochemical reduction and catalysis of cytochrome P450 3A4 in positive direction by 0.070-0.050 V, which indicates a thermodynamically more favorable process of electron transfer and enzymatic electrocatalysis.


Asunto(s)
Óxido de Aluminio , Nanoestructuras , Electrodos , Catálisis
3.
Biomed Khim ; 59(4): 378-87, 2013.
Artículo en Ruso | MEDLINE | ID: mdl-24502136

RESUMEN

Atomic force microscopy with two types of probes - standard (radius of curvature R approximately 10 nm) and supersharp (R approximately 2 nm)- was used to determine CYP102A1oligomeric state. CYP102A1 images were obtained in a liquid, air and vacuum environment using the standard probes, also a ratio of monomers to oligomers (alpha) of CYP102A1 were determined as alpha=0.48:0.52. At the same time use of standard probes did not allow to resolve the structure of these oligomers. Supersharp probes allowed to obtain the data about the monomers to oligomers ratio, and also about the dimers/trimers/tetramers ratio in air and vacuum. So, a ratio alpha of CYP102A1 in liquid can be determined by the standard probes, and an oligomeric state of protein can be specified by the supersharp probes.


Asunto(s)
Proteínas Bacterianas/química , Sistema Enzimático del Citocromo P-450/química , Microscopía de Fuerza Atómica , Complejos Multienzimáticos/química , Complejos Multienzimáticos/ultraestructura , NADPH-Ferrihemoproteína Reductasa/química , Estructura Cuaternaria de Proteína
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