Broad Neutralization Capacity of an Engineered Thermostable Three-Helix Angiotensin-Converting Enzyme 2 Polypeptide Targeting the Receptor-Binding Domain of SARS-CoV-2 DOI Open Access
Davide Cavazzini, Elisabetta Levati, S. Germani

и другие.

International Journal of Molecular Sciences, Год журнала: 2024, Номер 25(22), С. 12319 - 12319

Опубликована: Ноя. 16, 2024

The mutational drift of SARS-CoV-2 and the appearance multiple variants, including latest Omicron variant its sub-lineages, has significantly reduced (and in some cases abolished) protective efficacy Wuhan spike-antigen-based vaccines therapeutic antibodies. One most functionally constrained thus largely invariable regions spike protein is one involved interaction with ACE2 receptor mediating cellular entry SARS-CoV-2. Engineered ACE2, both as a full-length or an engineered polypeptide fragment, been shown to be capable preventing host-cell binding all viral variants endowed potent neutralization activity vitro vivo. Here, we report on biochemical antiviral properties rationally designed N-terminal, three-helix fragments that retain native-like conformation. these fragments, designated PRP8_3H produced recombinant form, bears structure-stabilizing binding-affinity enhancing mutations α-helix-I α-helix I II, respectively. While native-like, unmodified three α-helices fragment proved thermally unstable without any detectable pseudovirion capacity, was found highly thermostable receptor-binding domain nanomolar affinity neutralize spike-expressing pseudovirions at (sub)micromolar concentrations. lends itself promising decoy prototype suitable for variety formulations prophylactic applications.

Язык: Английский

Structure and Dynamics of Alpha B.1.1.7 SARS-CoV-2 S-Protein in complex with Fab of neutralizing antibody REGN10987 DOI
Milita V. Kocharovskaya, Evgeny Pichkur,

Artem D Ivannikov

и другие.

Biochemical and Biophysical Research Communications, Год журнала: 2025, Номер 755, С. 151558 - 151558

Опубликована: Фев. 27, 2025

Язык: Английский

Процитировано

0
Rapid In Vivo Screening of Monoclonal Antibody Cocktails Using Hydrodynamic Delivery of DNA-Encoded Modified Antibodies DOI Creative Commons

Hugues Fausther‐Bovendo,

George Babuadze,

Teodora Ivanciuc

и другие.

Biomedicines, Год журнала: 2025, Номер 13(3), С. 637 - 637

Опубликована: Март 5, 2025

Background: Monoclonal antibodies (mAbs) are potent treatment options for infectious diseases. The rapid isolation and in vivo validation of therapeutic mAb candidates, including cocktails, essential to combat novel or rapidly mutating pathogens. selection production candidates sufficient amount quality preclinical studies a major limiting step the development pipeline. Methods: Here, we developed method facilitate screening mAbs mouse models. Four conventional were transformed into single-chain variable fragments fused fragment crystallizable (Fc) region human IgG1 (scFv-IgG). These scFv-IgG expressed individually as cocktail vitro mice following transfection hydrodynamic delivery corresponding plasmids. Results: This induced high expression all provided protection two murine infection Conclusions: study highlights benefits this approach rapid, low-cost candidates.

Язык: Английский

Процитировано

0
An Improved Rapid and Sensitive Long Amplicon Method for Nanopore‐Based RSV Whole‐Genome Sequencing DOI Creative Commons
Xiaomin Dong, Steven W. Edwards, Yi‐Mo Deng

и другие.

Influenza and Other Respiratory Viruses, Год журнала: 2025, Номер 19(5)

Опубликована: Апрель 28, 2025

ABSTRACT Background Whole‐genome sequencing (WGS) provides critical insights into the respiratory syncytial virus (RSV) transmission and any emerging mutations that could impair efficacy of monoclonal antibodies or vaccines have been recently licenced for clinical use worldwide. However, ability to sequence RSV genomes at large scale is limited by expensive time‐consuming methods. Oxford Nanopore Technology (ONT) offers significant improvements in next generation (NGS) both turnaround time cost, compared with other platforms viral WGS. Methods We developed modified an long amplicon‐based WGS protocol ONT platform using a one‐step multiplex RT‐PCR assay rapid barcoding kit. One hundred thirty‐five positive Australian specimens (91 RSV‐A 44 RSV‐B) sampled 2023 cycle threshold (Ct) values between 14 35 were tested this study. This workflow was recent amplification assays based on short amplicons. Results A PCR amplicon clean‐up step prior library preparation significantly improved result samples poor generation, but it not necessary beneficial ones generated high concentrations Overall, success rate 85.9% achieved method performed as well more complex methods terms genome coverage depth. Conclusions The described here highly successful generating had times excellent results Ct up 30.

Язык: Английский

Процитировано

0
Structure and dynamics of the interaction of Delta and Omicron BA.1 SARS-CoV-2 variants with REGN10987 Fab reveal mechanism of antibody action DOI Creative Commons
Ekaterina N. Lyukmanova, Evgeny Pichkur, Dmitry E. Nolde

и другие.

Communications Biology, Год журнала: 2024, Номер 7(1)

Опубликована: Дек. 24, 2024

Study of mechanisms by which antibodies recognize different viral strains is necessary for the development new drugs and vaccines to treat COVID-19 other infections. Here, we report 2.5 Å cryo-EM structure SARS-CoV-2 Delta trimeric S-protein in complex with Fab recombinant analog REGN10987 neutralizing antibody. adopts "two RBD-down one RBD-up" conformation. interacts RBDs both conformations, blocking recognition angiotensin converting enzyme-2. Three-dimensional variability analysis reveals high mobility RBD/Fab regions. Interaction Wuhan, Delta, Omicron BA.1, mutated variants analyzed microscale thermophoresis, molecular dynamics simulations, ΔG calculations umbrella sampling one-dimensional potential mean force. Variability trajectories results a large scatter calculated values, but Boltzmann weighting provides an acceptable correlation experiment. evasion variant found be due additive effect N440K G446S mutations located at binding interface small Q498R mutation. Our study explains influence known-to-date RBD on highlights importance data beyond static complex. Cryo-EM Sprotein REGN10987, MD simulations variants, explain role individual evasion.

Язык: Английский

Процитировано

1
Broad Neutralization Capacity of an Engineered Thermostable Three-Helix Angiotensin-Converting Enzyme 2 Polypeptide Targeting the Receptor-Binding Domain of SARS-CoV-2 DOI Open Access
Davide Cavazzini, Elisabetta Levati, S. Germani

и другие.

International Journal of Molecular Sciences, Год журнала: 2024, Номер 25(22), С. 12319 - 12319

Опубликована: Ноя. 16, 2024

The mutational drift of SARS-CoV-2 and the appearance multiple variants, including latest Omicron variant its sub-lineages, has significantly reduced (and in some cases abolished) protective efficacy Wuhan spike-antigen-based vaccines therapeutic antibodies. One most functionally constrained thus largely invariable regions spike protein is one involved interaction with ACE2 receptor mediating cellular entry SARS-CoV-2. Engineered ACE2, both as a full-length or an engineered polypeptide fragment, been shown to be capable preventing host-cell binding all viral variants endowed potent neutralization activity vitro vivo. Here, we report on biochemical antiviral properties rationally designed N-terminal, three-helix fragments that retain native-like conformation. these fragments, designated PRP8_3H produced recombinant form, bears structure-stabilizing binding-affinity enhancing mutations α-helix-I α-helix I II, respectively. While native-like, unmodified three α-helices fragment proved thermally unstable without any detectable pseudovirion capacity, was found highly thermostable receptor-binding domain nanomolar affinity neutralize spike-expressing pseudovirions at (sub)micromolar concentrations. lends itself promising decoy prototype suitable for variety formulations prophylactic applications.

Язык: Английский

Процитировано

0