Biology subjects

Dzharullaeva, A. S.

Publications and source records attributed to Dzharullaeva, A. S..

3 recordsLinked to original sources

bioRxiv2025

Circular mRNA against Clean Cap linear mRNA vectors comprehensive comparison, expression, active and passive immunization

The mRNA platform has revolutionised vaccine technology by providing a universal, fast and easily scalable production process. There are two main types of mRNA - linear (cap-dependent) and circular (cap-independent), each with its own advantages. Although both vector types are continuously being improved, there has been no comprehensive comparative analysis of the most efficient existing vectors of each type. This paper provides a comparison of expression efficiency, protective and therapeutic activities of circular and linear mRNA vectors. We examined different combinations of linear vectors: containing either N1-methylpseudouridines or uridine and capped with either ARCA (m7G(5')ppp(5')G) or CleanCap (m7G(5')ppp(5')m2G). Circular vectors contained both commonly used IRES of coxsackievirus B3 and new a IRES of human rhinoviruses B6. Preliminary luciferase assay showed that modified linear vectors exhibited significantly higher expression levels both in vitro and in vivo. A similar but less dramatic difference was observed in expression of the target protein. At the same time, immunogenicity and protective activity of linear and circular vectors were equal.

immunology↗

bioRxiv2022

Single-domain antibody delivery using an mRNA platform protects against lethal doses of botulinum neurotoxin A

Single-domain antibodies (sdAbs, VHHs, or nanobodies) are a promising tool for the treatment of both infectious and somatic diseases. Their small size greatly simplifies any genetic engineering manipulations. Such antibodies have the ability to bind hard-to-reach antigenic epitopes through long parts of the variable chains, the third complementarity-determining regions (CDR3s). VHH fusion with the canonical immunoglobulin Fc fragment allows the Fc-fusion single-domain antibodies (VHH-Fc) to significantly increase their neutralizing activity and serum half-life. Previously we have developed and characterized VHH-Fc specific to botulinum neurotoxin A (BoNT/A), that showed a 1000-fold higher protective activity than monomeric form when challenged with five times the lethal dose (5 LD50) of BoNT/A. During the COVID-19 pandemic, mRNA vaccines based on lipid nanoparticles (LNP) as a delivery system have become an important translational technology that has significantly accelerated the clinical introduction of mRNA platforms. We have developed an mRNA platform that provides long-term expression after both intramuscular and intravenous application. The platform has been extensively characterized using firefly luciferase (Fluc) as a reporter. An intramuscular administration of LNP-mRNA encoding VHH-Fc antibody made it possible to achieve its rapid expression in mice and resulted in 100% protection when challenged with up to 100 LD50 of BoNT/A. The presented approach for the delivery of sdAbs using mRNA technology greatly simplifies drug development for antibody therapy and can be used for emergency prophylaxis.

molecular biology↗

bioRxiv2022

Cross-reactive single-domain antibodies to hemagglutinin stem region protect mice from group 1 influenza A virus infection

The continued evolution of influenza viruses reduces the effectiveness of vaccination and antiviral drugs. The identification of novel and universal agents for influenza prophylaxis and treatment is an urgent need. We have previously described two potent single-domain antibodies (VHH), G2.3 and H1.2, which efficiently neutralize H1N1 and H5N2 influenza viruses in vivo. In this study, we modified these VHHs with Fc-fragment to enhance their antiviral activity. Reformatting of G2.3 into bivalent Fc-fusion molecule increased its in vitro neutralizing activity against H1N1 and H2N3 viruses up to 20-fold and, moreover, resulted in obtaining the ability to neutralize H5N2 and H9N2 subtypes. We demonstrated that a dose as low as 0.6 mg/kg of G2.3-Fc or H1.2-Fc administered systemically or locally before infection could protect mice from lethal challenges with both H1N1 and H5N2 viruses. Furthermore, G2.3-Fc reduced the lung viral load to an undetectable level. Both VHH-Fc showed in vivo therapeutic efficacy when delivered via systemic or local route. The findings support G2.3-Fc as a potential therapeutic agent for both prophylaxis and therapy of Group 1 influenza A infection.

immunology↗