85 research outputs found
B1 plus exam : a key to higher education in Russia
The use of open electronic courses is a relatively new tendency in education and it is not yet fully discovered by researchers. This article examines the specifics of the introduction of online courses, including the requirements to them, as well as factors that slow down their active development.
The following methods of investigation were used: analysis, synthesis, deduction and induction. As an example the Kozma Minin Nizhny Novgorod State Pedagogical University - Minin University was used. On the basis of the study of the "Regulations on the use of open courses in the educational activities of Minin University", it was noted, in addition to the functions of the coordination group on open education, that for the trainee at the end of the course, in the case of unsatisfactory result, the possibility of the recalculation is established.
This possibility is used not by every university. In the article we make a conclusion that open courses are a new qualitative step in the development of world education, but online courses would be much more effective as narrow-minded, adult-oriented, and deeply motivated for learning. Taken as the basis of research, Minin University showed that the university is ready for the implementation of online courses. It is one of the few universities that actively use open courses in Russia.peer-reviewe
L-NAME-induced preeclampsia: correction of functional disorders of the hemostasis system with resveratrol and nicorandil
Preeclampsia is a formidable disease of the second half of pregnancy, leading to severe complications, including disability and even death. Many authors have recognized the correlation between the severity of preeclampsia and the degree of disturbances in the hemostasis system. In this regard, the objective of this study was to assess inhibition of platelet aggregation and the possibility of its correction with resverΠ°trol and nicorand
The influence of imidazoline receptor agonists on the ophthalmoscopic and morphometric parameters in retina in simulated retinal ischemia-reperfusion
An important task of experimental and clinical pharmacology and ophtalmology is to find highly effective drugs for correcting retinal ischemi
Search and evaluation of pharmacodynamic and pharmacokinetic parameters of selective blocker of TRPAβ ion channels from the group of substituted pyrazinopyrimidinones
Search considerations evaluation of pharmacodynamic and pharmacokinetic parameters of selective blocker of TRPAβ ion channels from the group of substituted pyrazinopyrimidinone
Review of a new concept of glaucoma pathogenesis based on the glymphatic theory of cerebrospinal fluid circulation
Clinical evidence for the glymphatic system hypothesis: There is some clinical, including case-based, and epidemiological evidence for similarities between glaucomatous optic nerve/retinal injuries and AD, since both occur in the form of improper secretion of neurotoxic metabolites, and both are often diagnosed togethe
ROLE OF MESENCHYMAL MULTIPOTENT STROMAL CELLS IN REMODELING OF BONE DEFECTS
Ability of mesenchymal multipotent stromal cells (MSCs) to differentiate into several types of mesenchymal tissues allows to consider these cells the main candidates for creating tissue engineeringΒ constructions for regenerative medicine. MSCs promote integration of bio-implants into the native bone and stimulate osteogenesis. MSCs are characterized by immunomodulatory properties, due to inflammation control and modification of immune cells. MSCs affect not only the in vivo immune response by preventing immunological rejection of implanted tissue engineering designs, but it can also influence the bone tissue immunity. MSCs play an important role in bone regeneration, by regulating the osteoblastic generation, and suppressing activity of inflammation effectors and osteoclastogenesis. Some pre-clinical and first clinical trials of bone bio-implants colonized with MSC, demonstrate promising outlooks for this strategy in order to obtain tissue engineering constructions for bone regeneration
Production and characterisation of a SARS-CoV-2 S-protein RBD homodimer with increased avidity for specific antibodies
Monitoring of the proportion of immune individuals and the effectiveness of vaccination in a population involves evaluation of several important parameters, including the level of virus-neutralising antibodies. In order to combat the COVID-19 pandemic, it is essential to develop approaches to detecting SARS-CoV-2 neutralising antibodies by safe, simple and rapid methods that do not require live viruses. To develop a test system for enzyme-linked immunosorbent assay (ELISA) that detects potential neutralising antibodies, it is necessary to obtain a highly purified recombinant receptor-binding domain (RBD) of the spike (S) protein with high avidity for specific antibodies.The aim of the study was to obtain and characterise a SARS-CoV-2 S-protein RBD homodimer and a recombinant RBD-expressing cell line, as well as to create an ELISA system for detecting potential neutralising antibodies.Materials and methods: the genetic construct was designed in silico. To generate a stable producer cell line, the authors transfected CHO-S cells, subjected them to antibiotic pressure, and selected the optimal clone. To isolate monomeric and homodimeric RBD forms, the authors purified the recombinant RBD by chromatographic methods. Further, they analysed the activity of the RBD forms by Western blotting, bio-layer interferometry, and indirect ELISA. The analysis involved mono clonal antibodies GamXRH19, GamP2C5, and h6g3, as well as serum samples from volunteers vaccinated with Gam-COVID-Vac (Sputnik V) and unvaccinated ones.Results: the authors produced the CHO-S cell line for stable expression of the recombinant SARS-CoV-2 S-protein RBD. The study demonstrated the recombinant RBDβs ability to homodimerise after fed-batch cultivation of the cell line for more than 7 days due to the presence of unpaired cysteines. The purified recombinant RBD yield from culture broth was 30β50 mg/L. Monomeric and homodimeric RBD forms were separated using gel-filtration chromatography and characterised by their ability to interact with specific monoclonal antibodies, as well as with serum samples from vaccinated volunteers. The homodimeric recombinant RBD showed increased avidity for both monoclonal and immune sera antibodies.Conclusions: the homodimeric recombinant RBD may be more preferable for the analysis of levels of antibodies to the receptor-binding domain of the SARS-CoV-2 S protein
Π Π²ΠΎΠΏΡΠΎΡΡ ΠΎ ΠΊΡΠΈΡΠΈΡΠ΅ΡΠΊΠΎΠΌ Π°ΡΡΠ΅ΡΠΈΠ°Π»ΡΠ½ΠΎΠΌ ΡΡΠ΅Π½ΠΎΠ·Π΅
Cardiovascular disease (CVD) is the leading cause of death worldwide. The main contribution to the structure of mortality from CVD is made by atherosclerosis. Indications for surgical treatment of patients with diseases caused by atherosclerotic lesions of the arteries are set taking into account the degree of their stenosis. Angiography has been considered the gold standard for screening patients with CVD for many years. Numerous studies carried out over the past several decades have revealed the weak side of this method in assessing the significance of borderline stenosis. Therefore, to analyze such changes, functional tests were introduced to clarify the indications for surgical intervention. Currently, criteria for the significance of stenosis of the iliac, renal and coronary arteries have been determined. The significance of stenosis of the arteries supplying the brain and intestines is still a matter of debate and requires further study.Π‘Π΅ΡΠ΄Π΅ΡΠ½ΠΎ-ΡΠΎΡΡΠ΄ΠΈΡΡΡΠ΅ Π·Π°Π±ΠΎΠ»Π΅Π²Π°Π½ΠΈΡ (Π‘Π‘Π) Π·Π°Π½ΠΈΠΌΠ°ΡΡ 1-Π΅ ΠΌΠ΅ΡΡΠΎ ΡΡΠ΅Π΄ΠΈ ΠΏΡΠΈΡΠΈΠ½ ΡΠΌΠ΅ΡΡΠΈ Π²ΠΎ Π²ΡΠ΅ΠΌ ΠΌΠΈΡΠ΅. ΠΡΠ½ΠΎΠ²Π½ΠΎΠΉ Π²ΠΊΠ»Π°Π΄ Π² ΡΡΡΡΠΊΡΡΡΡ ΡΠΌΠ΅ΡΡΠ½ΠΎΡΡΠΈ ΠΎΡ Π‘Π‘Π Π²Π½ΠΎΡΠΈΡ Π°ΡΠ΅ΡΠΎΡΠΊΠ»Π΅ΡΠΎΠ·. ΠΠΎΠΊΠ°Π·Π°Π½ΠΈΡ ΠΊ Ρ
ΠΈΡΡΡΠ³ΠΈΡΠ΅ΡΠΊΠΎΠΌΡ Π»Π΅ΡΠ΅Π½ΠΈΡ ΠΏΠ°ΡΠΈΠ΅Π½ΡΠΎΠ² Ρ Π·Π°Π±ΠΎΠ»Π΅Π²Π°Π½ΠΈΡΠΌΠΈ, Π²ΡΠ·Π²Π°Π½Π½ΡΠΌΠΈ Π°ΡΠ΅ΡΠΎΡΠΊΠ»Π΅ΡΠΎΡΠΈΡΠ΅ΡΠΊΠΈΠΌ ΡΡΠΆΠ΅Π½ΠΈΠ΅ΠΌ Π°ΡΡΠ΅ΡΠΈΠΉ, Π²ΡΡΡΠ°Π²Π»ΡΡΡΡΡ Ρ ΡΡΠ΅ΡΠΎΠΌ ΡΡΠ΅ΠΏΠ΅Π½ΠΈ ΠΈΡ
ΡΡΠ΅Π½ΠΎΠ·Π°. ΠΠ° ΠΏΡΠΎΡΡΠΆΠ΅Π½ΠΈΠΈ ΠΌΠ½ΠΎΠ³ΠΈΡ
Π»Π΅Ρ Β«Π·ΠΎΠ»ΠΎΡΡΠΌ ΡΡΠ°Π½Π΄Π°ΡΡΠΎΠΌΒ» ΠΎΠ±ΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ ΠΏΠ°ΡΠΈΠ΅Π½ΡΠΎΠ² Ρ Π‘Π‘Π ΡΡΠΈΡΠ°Π΅ΡΡΡ Π°Π½Π³ΠΈΠΎΠ³ΡΠ°ΡΠΈΡ. ΠΠ½ΠΎΠΆΠ΅ΡΡΠ²Π΅Π½Π½ΡΠ΅ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ, ΠΏΡΠΎΠ²Π΅Π΄Π΅Π½Π½ΡΠ΅ Π·Π° ΠΏΠΎΡΠ»Π΅Π΄Π½ΠΈΠ΅ Π½Π΅ΡΠΊΠΎΠ»ΡΠΊΠΎ Π΄Π΅ΡΡΡΠΊΠΎΠ² Π»Π΅Ρ, Π²ΡΡΠ²ΠΈΠ»ΠΈ ΡΠ»Π°Π±ΡΡ ΡΡΠΎΡΠΎΠ½Ρ ΡΡΠΎΠ³ΠΎ ΠΌΠ΅ΡΠΎΠ΄Π° Π² ΠΎΡΠ΅Π½ΠΊΠ΅ Π·Π½Π°ΡΠΈΠΌΠΎΡΡΠΈ ΠΏΠΎΠ³ΡΠ°Π½ΠΈΡΠ½ΡΡ
ΡΡΠ΅Π½ΠΎΠ·ΠΎΠ². ΠΠΎΡΡΠΎΠΌΡ Π΄Π»Ρ Π°Π½Π°Π»ΠΈΠ·Π° ΡΠ°ΠΊΠΈΡ
ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΠΉ Π±ΡΠ»ΠΈ Π²Π²Π΅Π΄Π΅Π½Ρ ΡΡΠ½ΠΊΡΠΈΠΎΠ½Π°Π»ΡΠ½ΡΠ΅ ΡΠ΅ΡΡΡ, ΡΡΠΎΡΠ½ΡΡΡΠΈΠ΅ ΠΏΠΎΠΊΠ°Π·Π°Π½ΠΈΡ ΠΊ Ρ
ΠΈΡΡΡΠ³ΠΈΡΠ΅ΡΠΊΠΎΠΌΡ Π²ΠΌΠ΅ΡΠ°ΡΠ΅Π»ΡΡΡΠ²Ρ. Π Π½Π°ΡΡΠΎΡΡΠ΅Π΅ Π²ΡΠ΅ΠΌΡ ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½Ρ ΠΊΡΠΈΡΠ΅ΡΠΈΠΈ Π·Π½Π°ΡΠΈΠΌΠΎΡΡΠΈ ΡΡΠΆΠ΅Π½ΠΈΡ ΠΏΠΎΠ΄Π²Π·Π΄ΠΎΡΠ½ΡΡ
, ΠΏΠΎΡΠ΅ΡΠ½ΡΡ
ΠΈ ΠΊΠΎΡΠΎΠ½Π°ΡΠ½ΡΡ
Π°ΡΡΠ΅ΡΠΈΠΉ. ΠΠ½Π°ΡΠΈΠΌΠΎΡΡΡ ΡΡΠ΅Π½ΠΎΠ·ΠΎΠ² Π°ΡΡΠ΅ΡΠΈΠΉ, ΠΊΡΠΎΠ²ΠΎΡΠ½Π°Π±ΠΆΠ°ΡΡΠΈΡ
Π³ΠΎΠ»ΠΎΠ²Π½ΠΎΠΉ ΠΌΠΎΠ·Π³ ΠΈ ΠΊΠΈΡΠ΅ΡΠ½ΠΈΠΊ, Π΄ΠΎ ΡΠΈΡ
ΠΏΠΎΡ ΠΎΡΡΠ°Π΅ΡΡΡ ΠΏΡΠ΅Π΄ΠΌΠ΅ΡΠΎΠΌ Π΄ΠΈΡΠΊΡΡΡΠΈΠΉ ΠΈ ΡΡΠ΅Π±ΡΠ΅Ρ Π΄Π°Π»ΡΠ½Π΅ΠΉΡΠ΅Π³ΠΎ ΠΈΠ·ΡΡΠ΅Π½ΠΈΡ
ΠΠΎΠ»ΡΡΠ΅Π½ΠΈΠ΅ ΠΈ Ρ Π°ΡΠ°ΠΊΡΠ΅ΡΠΈΡΡΠΈΠΊΠ° Π³ΠΎΠΌΠΎΠ΄ΠΈΠΌΠ΅ΡΠ½ΠΎΠΉ ΡΠΎΡΠΌΡ RBD S-Π±Π΅Π»ΠΊΠ° SARS-CoV-2, ΠΎΠ±Π»Π°Π΄Π°ΡΡΠ΅ΠΉ ΠΏΠΎΠ²ΡΡΠ΅Π½Π½ΠΎΠΉ Π°Π²ΠΈΠ΄Π½ΠΎΡΡΡΡ ΠΊ ΡΠΏΠ΅ΡΠΈΡΠΈΡΠ΅ΡΠΊΠΈΠΌ Π°Π½ΡΠΈΡΠ΅Π»Π°ΠΌ
Monitoring of the proportion of immune individuals and the effectiveness of vaccination in a population involves evaluation of several important parameters, including the level of virus-neutralising antibodies. In order to combat the COVID-19 pandemic, it is essential to develop approaches to detecting SARS-CoV-2 neutralising antibodies by safe, simple and rapid methods that do not require live viruses. To develop a test system for enzyme-linked immunosorbent assay (ELISA) that detects potential neutralising antibodies, it is necessary to obtain a highly purified recombinant receptor-binding domain (RBD) of the spike (S) protein with high avidity for specific antibodies. The aim of the study w as t o obtain and characterise a SARSCoV-2 S-protein RBD homodimer and a recombinant RBD-expressing cell line, as well as to create an ELISA system for detecting potential neutralising antibodies. Materials and methods: the genetic construct was designed in silico. To generate a stable producer cell line, the authors transfected CHO-S cells, subjected them to antibiotic pressure, and selected the optimal clone. To isolate monomeric and homodimeric RBD forms, the authors purified the recombinant RBD by chromatographic methods. Further, they analysed the activity of the RBD forms by Western blotting, bio-layer interferometry, and indirect ELISA. The analysis involved monoclonal antibodies GamXRH19, GamP2C5, and h6g3, as well as serum samples from volunteers vaccinated with Gam-COVID-Vac (Sputnik V) and unvaccinated ones. Results: the authors produced the CHO-S cell line for stable expression of the recombinant SARS-CoV-2 S-protein RBD. The study demonstrated the recombinant RBDβs ability to homodimerise after fed-batch cultivation of the cell line for more than 7 days due to the presence of unpaired cysteines. The purified recombinant RBD yield from culture broth was 30β50 mg/L. Monomeric and homodimeric RBD forms were separated using gel-filtration chromatography and characterised by their ability to interact with specific monoclonal antibodies, as well as with serum samples from vaccinated volunteers. The homodimeric recombinant RBD showed increased avidity for both monoclonal and immune sera antibodies. Conclusions: the homodimeric recombinant RBD may be more preferable for the analysis of levels of antibodies to the receptor-binding domain of the SARS-CoV-2 S protein.ΠΠ°ΠΆΠ½ΡΠΌ ΠΏΠ°ΡΠ°ΠΌΠ΅ΡΡΠΎΠΌ, ΠΎΡΠ΅Π½ΠΈΠ²Π°Π΅ΠΌΡΠΌ ΠΏΡΠΈ ΠΌΠΎΠ½ΠΈΡΠΎΡΠΈΠ½Π³Π΅ ΠΈΠΌΠΌΡΠ½Π½ΠΎΠΉ ΠΏΡΠΎΡΠ»ΠΎΠΉΠΊΠΈ Ρ Π½Π°ΡΠ΅Π»Π΅Π½ΠΈΡ ΠΈ ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ Π²Π°ΠΊΡΠΈΠ½Π°ΡΠΈΠΈ Π½Π°ΡΠ΅Π»Π΅Π½ΠΈΡ, ΡΠ²Π»ΡΠ΅ΡΡΡ ΡΡΠΎΠ²Π΅Π½Ρ Π²ΠΈΡΡΡΠ½Π΅ΠΉΡΡΠ°Π»ΠΈΠ·ΡΡΡΠΈΡ
Π°Π½ΡΠΈΡΠ΅Π». Π Π°Π·ΡΠ°Π±ΠΎΡΠΊΠ° ΠΏΠΎΠ΄Ρ
ΠΎΠ΄Π° ΠΊ Π²ΡΡΠ²Π»Π΅Π½ΠΈΡ Π²ΠΈΡΡΡΠ½Π΅ΠΉΡΡΠ°Π»ΠΈΠ·ΡΡΡΠΈΡ
Π°Π½ΡΠΈΡΠ΅Π» ΠΊ Π²ΠΈΡΡΡΡ SARS-CoV-2 Ρ ΠΏΠΎΠΌΠΎΡΡΡ Π±Π΅Π·ΠΎΠΏΠ°ΡΠ½ΠΎΠ³ΠΎ, ΠΏΡΠΎΡΡΠΎΠ³ΠΎ ΠΈ Π±ΡΡΡΡΠΎΠ³ΠΎ ΠΌΠ΅ΡΠΎΠ΄Π°, Π½Π΅ ΡΡΠ΅Π±ΡΡΡΠ΅Π³ΠΎ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΡ ΠΆΠΈΠ²ΡΡ
Π²ΠΈΡΡΡΠΎΠ², ΠΈΠΌΠ΅Π΅Ρ Π±ΠΎΠ»ΡΡΠΎΠ΅ Π·Π½Π°ΡΠ΅Π½ΠΈΠ΅ Π΄Π»Ρ Π±ΠΎΡΡΠ±Ρ Ρ ΠΏΠ°Π½Π΄Π΅ΠΌΠΈΠ΅ΠΉ COVID-19. ΠΠ»Ρ ΡΠ°Π·ΡΠ°Π±ΠΎΡΠΊΠΈ ΡΠ΅ΡΡ-ΡΠΈΡΡΠ΅ΠΌ Π΄Π»Ρ ΠΏΡΠΎΠ²Π΅Π΄Π΅Π½ΠΈΡ ΠΈΠΌΠΌΡΠ½ΠΎΡΠ΅ΡΠΌΠ΅Π½ΡΠ½ΠΎΠ³ΠΎ Π°Π½Π°Π»ΠΈΠ·Π° (ΠΠ€Π), Π΄Π΅ΡΠ΅ΠΊΡΠΈΡΡΡΡΠΈΡ
ΠΏΠΎΡΠ΅Π½ΡΠΈΠ°Π»ΡΠ½ΠΎ Π²ΠΈΡΡΡΠ½Π΅ΠΉΡΡΠ°Π»ΠΈΠ·ΡΡΡΠΈΠ΅ Π°Π½ΡΠΈΡΠ΅Π»Π°, Π½Π΅ΠΎΠ±Ρ
ΠΎΠ΄ΠΈΠΌΠΎ ΠΏΠΎΠ»ΡΡΠ΅Π½ΠΈΠ΅ Π²ΡΡΠΎΠΊΠΎΠΎΡΠΈΡΠ΅Π½Π½ΠΎΠ³ΠΎ ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΠΎΠ³ΠΎ ΡΠ΅ΡΠ΅ΠΏΡΠΎΡ-ΡΠ²ΡΠ·ΡΠ²Π°ΡΡΠ΅Π³ΠΎ Π΄ΠΎΠΌΠ΅Π½Π° (RBD) S-Π±Π΅Π»ΠΊΠ°, ΠΎΠ±Π»Π°Π΄Π°ΡΡΠ΅Π³ΠΎ Π²ΡΡΠΎΠΊΠΎΠΉ Π°Π²ΠΈΠ΄Π½ΠΎΡΡΡΡ ΠΊ ΡΠΏΠ΅ΡΠΈΡΠΈΡΠ΅ΡΠΊΠΈΠΌ Π°Π½ΡΠΈΡΠ΅Π»Π°ΠΌ. Π¦Π΅Π»Ρ ΡΠ°Π±ΠΎΡΡ: ΠΏΠΎΠ»ΡΡΠ΅Π½ΠΈΠ΅ ΠΈ Ρ
Π°ΡΠ°ΠΊΡΠ΅ΡΠΈΡΡΠΈΠΊΠ° Π³ΠΎΠΌΠΎΠ΄ΠΈΠΌΠ΅ΡΠ½ΠΎΠΉ ΡΠΎΡΠΌΡ RBD S-Π±Π΅Π»ΠΊΠ° Π²ΠΈΡΡΡΠ° SARS-CoV-2, Π° ΡΠ°ΠΊΠΆΠ΅ ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠΉ Π»ΠΈΠ½ΠΈΠΈ, ΠΏΡΠΎΠ΄ΡΡΠΈΡΡΡΡΠ΅ΠΉ ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΡΠΉ RBD, Π΄Π»Ρ ΡΠΎΠ·Π΄Π°Π½ΠΈΡ ΠΠ€Π ΡΠ΅ΡΡ-ΡΠΈΡΡΠ΅ΠΌΡ Π΄Π»Ρ Π²ΡΡΠ²Π»Π΅Π½ΠΈΡ ΠΏΠΎΡΠ΅Π½ΡΠΈΠ°Π»ΡΠ½ΠΎ Π²ΠΈΡΡΡΠ½Π΅ΠΉΡΡΠ°Π»ΠΈΠ·ΡΡΡΠΈΡ
Π°Π½ΡΠΈΡΠ΅Π». ΠΠ°ΡΠ΅ΡΠΈΠ°Π»Ρ ΠΈ ΠΌΠ΅ΡΠΎΠ΄Ρ: Π΄ΠΈΠ·Π°ΠΉΠ½ Π³Π΅Π½Π΅ΡΠΈΡΠ΅ΡΠΊΠΎΠΉ ΠΊΠΎΠ½ΡΡΡΡΠΊΡΠΈΠΈ ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΠ»ΠΈ in silico. Π‘ΡΠ°Π±ΠΈΠ»ΡΠ½ΡΡ ΠΊΠ»Π΅ΡΠΎΡΠ½ΡΡ Π»ΠΈΠ½ΠΈΡ ΠΏΠΎΠ»ΡΡΠ°Π»ΠΈ ΠΏΡΠΈ ΠΏΠΎΠΌΠΎΡΠΈ ΡΡΠ°Π½ΡΡΠ΅ΠΊΡΠΈΠΈ ΠΊΠ»Π΅ΡΠΎΠΊ CHO-S, ΡΠ΅Π»Π΅ΠΊΡΠΈΠΈ Π½Π° Π°Π½ΡΠΈΠ±ΠΈΠΎΡΠΈΠΊΠ΅ ΠΈ ΠΎΡΠ±ΠΎΡΠ° ΠΎΠΏΡΠΈΠΌΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΠΊΠ»ΠΎΠ½Π°. Π Π΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΡΠΉ RBD ΠΎΡΠΈΡΠ°Π»ΠΈ Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ Ρ
ΡΠΎΠΌΠ°ΡΠΎΠ³ΡΠ°ΡΠΈΡΠ΅ΡΠΊΠΈΡ
ΠΌΠ΅ΡΠΎΠ΄ΠΎΠ², ΠΏΠΎΠ»ΡΡΠ°Π»ΠΈ ΠΌΠΎΠ½ΠΎΠΌΠ΅ΡΠ½ΡΡ ΠΈ Π³ΠΎΠΌΠΎΠ΄ΠΈΠΌΠ΅ΡΠ½ΡΡ ΡΠΎΡΠΌΡ RBD. ΠΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΠΏΠΎΠ»ΡΡΠ΅Π½Π½ΡΡ
ΡΠΎΡΠΌ Π°Π½Π°Π»ΠΈΠ·ΠΈΡΠΎΠ²Π°Π»ΠΈ Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠ² ΠΠ΅ΡΡΠ΅ΡΠ½-Π±Π»ΠΎΡ, Π±ΠΈΠΎΡΠ»ΠΎΠΉΠ½ΠΎΠΉ ΠΈΠ½ΡΠ΅ΡΡΠ΅ΡΠΎΠΌΠ΅ΡΡΠΈΠΈ ΠΈ Π½Π΅ΠΏΡΡΠΌΠ³ΠΎ ΠΠ€Π. ΠΠ»Ρ Π°Π½Π°Π»ΠΈΠ·Π° ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π»ΠΈ ΠΌΠΎΠ½ΠΎΠΊΠ»ΠΎΠ½Π°Π»ΡΠ½ΡΠ΅ Π°Π½ΡΠΈΡΠ΅Π»Π° GamXRH19, GamP2C5 ΠΈ h6g3, Π° ΡΠ°ΠΊΠΆΠ΅ ΠΎΠ±ΡΠ°Π·ΡΡ ΡΡΠ²ΠΎΡΠΎΡΠΎΠΊ ΠΊΡΠΎΠ²ΠΈ Π΄ΠΎΠ±ΡΠΎΠ²ΠΎΠ»ΡΡΠ΅Π², Π²Π°ΠΊΡΠΈΠ½ΠΈΡΠΎΠ²Π°Π½Π½ΡΡ
ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠΎΠΌ ΠΠ°ΠΌ-ΠΠΠΠΠ-ΠΠ°ΠΊ, ΠΈ Π½Π΅Π²Π°ΠΊΡΠΈΠ½ΠΈΡΠΎΠ²Π°Π½Π½ΡΡ
Π΄ΠΎΠ±ΡΠΎΠ²ΠΎΠ»ΡΡΠ΅Π². Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ: ΠΏΠΎΠ»ΡΡΠ΅Π½Π° ΠΊΠ»Π΅ΡΠΎΡΠ½Π°Ρ Π»ΠΈΠ½ΠΈΡ CHO-S, ΡΡΠ°Π±ΠΈΠ»ΡΠ½ΠΎ ΠΏΡΠΎΠ΄ΡΡΠΈΡΡΡΡΠ°Ρ ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΡΠΉ RBD S-Π±Π΅Π»ΠΊΠ° Π²ΠΈΡΡΡΠ° SARS-CoV-2. ΠΠΎΠΊΠ°Π·Π°Π½ΠΎ, ΡΡΠΎ ΠΏΡΠΈ ΠΊΡΠ»ΡΡΠΈΠ²ΠΈΡΠΎΠ²Π°Π½ΠΈΠΈ Π΄Π°Π½Π½ΠΎΠΉ ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠΉ Π»ΠΈΠ½ΠΈΠΈ Π² ΡΠ΅ΠΆΠΈΠΌΠ΅ fed-batch Π±ΠΎΠ»Π΅Π΅ 7 ΡΡΡΠΎΠΊ ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΡΠΉ RBD ΡΠΏΠΎΡΠΎΠ±Π΅Π½ ΠΎΠ±ΡΠ°Π·ΠΎΠ²ΡΠ²Π°ΡΡ Π³ΠΎΠΌΠΎΠ΄ΠΈΠΌΠ΅ΡΡ Π·Π° ΡΡΠ΅Ρ Π½Π°Π»ΠΈΡΠΈΡ Π½Π΅ΡΠΏΠ°ΡΠ΅Π½Π½ΡΡ
ΡΠΈΡΡΠ΅ΠΈΠ½ΠΎΠ². ΠΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π΅Π½Π½ΡΠΉ Π²ΡΡ
ΠΎΠ΄ ΠΎΡΠΈΡΠ΅Π½Π½ΠΎΠ³ΠΎ ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΠΎΠ³ΠΎ RBD ΠΈΠ· ΠΊΡΠ»ΡΡΡΡΠ°Π»ΡΠ½ΠΎΠΉ ΠΆΠΈΠ΄ΠΊΠΎΡΡΠΈ ΡΠΎΡΡΠ°Π²ΠΈΠ» 30β50 ΠΌΠ³/Π». ΠΠΎΠ½ΠΎΠΌΠ΅ΡΠ½Π°Ρ ΠΈ Π³ΠΎΠΌΠΎΠ΄ΠΈΠΌΠ΅ΡΠ½Π°Ρ ΡΠΎΡΠΌΡ RBD Π±ΡΠ»ΠΈ ΡΠ°Π·Π΄Π΅Π»Π΅Π½Ρ ΠΏΡΠΈ ΠΏΠΎΠΌΠΎΡΠΈ Π³Π΅Π»Ρ-ΡΠΈΠ»ΡΡΡΠ°ΡΠΈΠΈ ΠΈ ΠΎΡ
Π°ΡΠ°ΠΊΡΠ΅ΡΠΈΠ·ΠΎΠ²Π°Π½Ρ ΠΏΠΎ ΡΠΏΠΎΡΠΎΠ±Π½ΠΎΡΡΠΈ Π²Π·Π°ΠΈΠΌΠΎΠ΄Π΅ΠΉΡΡΠ²ΠΎΠ²Π°ΡΡ ΡΠΎ ΡΠΏΠ΅ΡΠΈΡΠΈΡΠ΅ΡΠΊΠΈΠΌΠΈ ΠΌΠΎΠ½ΠΎΠΊΠ»ΠΎΠ½Π°Π»ΡΠ½ΡΠΌΠΈ Π°Π½ΡΠΈΡΠ΅Π»Π°ΠΌΠΈ, Π° ΡΠ°ΠΊΠΆΠ΅ ΡΡΠ²ΠΎΡΠΎΡΠΊΠ°ΠΌΠΈ ΠΊΡΠΎΠ²ΠΈ ΠΎΡ Π²Π°ΠΊΡΠΈΠ½ΠΈΡΠΎΠ²Π°Π½Π½ΡΡ
Π΄ΠΎΠ±ΡΠΎΠ²ΠΎΠ»ΡΡΠ΅Π². ΠΡΠΎΠ΄Π΅ΠΌΠΎΠ½ΡΡΡΠΈΡΠΎΠ²Π°Π½ΠΎ, ΡΡΠΎ ΠΈΠΌΠ΅Π½Π½ΠΎ Π³ΠΎΠΌΠΎΠ΄ΠΈΠΌΠ΅ΡΠ½Π°Ρ ΡΠΎΡΠΌΠ° ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΠΎΠ³ΠΎ RBD ΠΎΠ±Π»Π°Π΄Π°Π΅Ρ ΠΏΠΎΠ²ΡΡΠ΅Π½Π½ΠΎΠΉ Π°Π²ΠΈΠ΄Π½ΠΎΡΡΡΡ ΠΊ ΠΌΠΎΠ½ΠΎΠΊΠ»ΠΎΠ½Π°Π»ΡΠ½ΡΠΌ Π°Π½ΡΠΈΡΠ΅Π»Π°ΠΌ ΠΈ Π°Π½ΡΠΈΡΠ΅Π»Π°ΠΌ Π² ΡΡΠ²ΠΎΡΠΎΡΠΊΠ΅ ΠΊΡΠΎΠ²ΠΈ Π²Π°ΠΊΡΠΈΠ½ΠΈΡΠΎΠ²Π°Π½Π½ΡΡ
. ΠΡΠ²ΠΎΠ΄Ρ: Π³ΠΎΠΌΠΎΠ΄ΠΈΠΌΠ΅ΡΠ½Π°Ρ ΡΠΎΡΠΌΠ° ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΠΎΠ³ΠΎ RBD ΠΌΠΎΠΆΠ΅Ρ ΡΠ²Π»ΡΡΡΡΡ Π±ΠΎΠ»Π΅Π΅ ΠΏΡΠ΅Π΄ΠΏΠΎΡΡΠΈΡΠ΅Π»ΡΠ½ΠΎΠΉ Π΄Π»Ρ Π°Π½Π°Π»ΠΈΠ·Π° ΡΡΠΎΠ²Π½Ρ Π°Π½ΡΠΈΡΠ΅Π» ΠΊ ΡΠ΅ΡΠ΅ΠΏΡΠΎΡ-ΡΠ²ΡΠ·ΡΠ²Π°ΡΡΠ΅ΠΌΡ Π΄ΠΎΠΌΠ΅Π½Ρ S-Π±Π΅Π»ΠΊΠ° Π²ΠΈΡΡΡΠ° SARS-CoV-2
rAAV expressing recombinant antibody for emergency prevention and long-term prophylaxis of COVID-19
IntroductionNumerous agents for prophylaxis of SARS-CoV-2-induced diseases are currently registered for the clinical use. Formation of the immunity happens within several weeks following vaccine administration which is their key disadvantage. In contrast, drugs based on monoclonal antibodies, enable rapid passive immunization and therefore can be used for emergency pre- and post-exposure prophylaxis of COVID-19. However rapid elimination of antibody-based drugs from the circulation limits their usage for prolonged pre-exposure prophylaxis.MethodsIn current work we developed a recombinant adeno-associated viral vector (rAAV), expressing a SARS-CoV-2 spike receptor-binding domain (RBD)-specific antibody P2C5 fused with a human IgG1 Fc fragment (P2C5-Fc) using methods of molecular biotechnology and bioprocessing.Results and discussionsA P2C5-Fc antibody expressed by a proposed rAAV (rAAV-P2C5-Fc) was shown to circulate within more than 300 days in blood of transduced mice and protect animals from lethal SARS-CoV-2 virus (B.1.1.1 and Omicron BA.5 variants) lethal dose of 105 TCID50. In addition, rAAV-P2C5-Fc demonstrated 100% protective activity as emergency prevention and long-term prophylaxis, respectively. It was also demonstrated that high titers of neutralizing antibodies to the SARS-CoV-2 virus were detected in the blood serum of animals that received rAAV-P2C5-Fc for more than 10 months from the moment of administration.Our data therefore indicate applicability of an rAAV for passive immunization and induction of a rapid long-term protection against various SARS-CoV-2 variants
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