A pseudovirus system enables deep mutational scanning of the full SARS-CoV-2 spike

Cell, Год журнала: 2023, Номер 186(6), С. 1263 - 1278.e20

Опубликована: Фев. 13, 2023

A major challenge in understanding SARS-CoV-2 evolution is interpreting the antigenic and functional effects of emerging mutations viral spike protein. Here, we describe a deep mutational scanning platform based on non-replicative pseudotyped lentiviruses that directly quantifies how large numbers impact antibody neutralization pseudovirus infection. We apply this to produce libraries Omicron BA.1 Delta spikes. These each contain ∼7,000 distinct amino acid context up ∼135,000 unique mutation combinations. use these map escape from neutralizing antibodies targeting receptor-binding domain, N-terminal S2 subunit spike. Overall, work establishes high-throughput safe approach measure ∼10

Язык: Английский

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Mapping SARS-CoV-2 antigenic relationships and serological responses

Science, Год журнала: 2023, Номер 382(6666)

Опубликована: Окт. 6, 2023

During the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, multiple variants escaping preexisting immunity emerged, causing reinfections of previously exposed individuals. Here, we used antigenic cartography to analyze patterns cross-reactivity among 21 and 15 groups human sera obtained after primary infection with 10 different or messenger RNA (mRNA)–1273 mRNA-1273.351 vaccination. We found differences pre-Omicron caused by substitutions at spike-protein positions 417, 452, 484, 501. Quantifying changes in response breadth over time additional vaccine doses, our results show largest increase between 4 weeks >3 months a second dose. immunodominance spike regions, depending on variant an individual was first to, implications for risk assessment vaccine-strain selection.

Язык: Английский

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Spike deep mutational scanning helps predict success of SARS-CoV-2 clades

Nature, Год журнала: 2024, Номер 631(8021), С. 617 - 626

Опубликована: Июль 3, 2024

SARS-CoV-2 variants acquire mutations in the spike protein that promote immune evasion

Язык: Английский

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Full-spike deep mutational scanning helps predict the evolutionary success of SARS-CoV-2 clades

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2023, Номер unknown

Опубликована: Ноя. 14, 2023

SARS-CoV-2 variants acquire mutations in spike that promote immune evasion and impact other properties contribute to viral fitness such as ACE2 receptor binding cell entry. Knowledge of how affect these phenotypes can provide insight into the current potential future evolution virus. Here we use pseudovirus deep mutational scanning measure >9,000 across full XBB.1.5 BA.2 spikes binding, entry, or escape from human sera. We find outside receptor-binding domain (RBD) have meaningfully impacted during evolution. also neutralization by serum individuals who recently had infections. The strongest are RBD at sites 357, 420, 440, 456, 473-however, antigenic impacts vary individuals. identify strong RBD; however many them decrease suggesting they act modulating conformation. Notably, growth rates clades be explained substantial part measured effects on phenotypes, our data could enable better prediction

Язык: Английский

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Antibodies targeting a quaternary site on SARS-CoV-2 spike glycoprotein prevent viral receptor engagement by conformational locking

Immunity, Год журнала: 2023, Номер 56(10), С. 2442 - 2455.e8

Опубликована: Сен. 29, 2023

SARS-CoV-2 continues to evolve, with many variants evading clinically authorized antibodies. To isolate monoclonal antibodies (mAbs) broadly neutralizing capacities against the virus, we screened serum samples from convalescing COVID-19 patients. We isolated two mAbs, 12-16 and 12-19, which neutralized all tested, including XBB subvariants, prevented infection in hamsters challenged Omicron BA.1 intranasally. Structurally, both targeted a conserved quaternary epitope located at interface between N-terminal domain subdomain 1, uncovering site of vulnerability on spike. These viral receptor engagement by locking receptor-binding (RBD) spike down conformation, revealing mechanism virus neutralization for non-RBD Deep mutational scanning showed that could mutate escape but such mutations are rarely found circulating viruses. Antibodies 12-19 hold promise as prophylactic agents immunocompromised persons who do not respond robustly vaccines.

Язык: Английский

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Research progress in methods for detecting neutralizing antibodies against SARS-CoV-2

Analytical Biochemistry, Год журнала: 2023, Номер 673, С. 115199 - 115199

Опубликована: Май 29, 2023

Язык: Английский

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Broad immunity to SARS-CoV-2 variants of concern mediated by a SARS-CoV-2 receptor-binding domain protein vaccine

EBioMedicine, Год журнала: 2023, Номер 92, С. 104574 - 104574

Опубликована: Май 4, 2023

The SARS-CoV-2 global pandemic has fuelled the generation of vaccines at an unprecedented pace and scale. However, many challenges remain, including: emergence vaccine-resistant mutant viruses, vaccine stability during storage transport, waning vaccine-induced immunity, concerns about infrequent adverse events associated with existing vaccines.We report on a protein subunit comprising receptor-binding domain (RBD) ancestral spike protein, dimerised immunoglobulin IgG1 Fc domain. These were tested in conjunction three different adjuvants: TLR2 agonist R4-Pam2Cys, NKT cell glycolipid α-Galactosylceramide, or MF59® squalene oil-in-water adjuvant, using mice, rats hamsters. We also developed RBD-human RBD sequence immuno-evasive beta variant (N501Y, E484K, K417N). as heterologous third dose booster following priming whole vaccine.Each formulation RBD-Fc drove strong neutralising antibody (nAb) responses provided durable highly protective immunity against lower upper airway infection mouse models COVID-19. 'beta variant' vaccine, combined induced protection mice strain well strain. Furthermore, when used booster, increased titres nAb other variants including alpha, delta, delta+, gamma, lambda, mu, omicron BA.1, BA.2 BA.5.These results demonstrated that subunit/MF59® adjuvanted can induce high levels broadly reactive nAbs, prior immunisation ancestral-strain vaccines. This platform offers potential approach to augment some currently approved face emerging concern, it now entered phase I clinical trial.This work was supported by grants from Medical Research Future Fund (MRFF) (2005846), Jack Ma Foundation, National Health Council Australia (NHMRC; 1113293) Singapore (MOH-COVID19RF-003). Individual researchers NHMRC Senior Principal Fellowship (1117766), Investigator Awards (2008913 1173871), Australian Discovery Early Career Award (ARC DECRA; DE210100705) philanthropic awards IFM investors A2 Milk Company.

Язык: Английский

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Uses and Challenges of Antiviral Polyclonal and Monoclonal Antibody Therapies

Pharmaceutics, Год журнала: 2023, Номер 15(5), С. 1538 - 1538

Опубликована: Май 19, 2023

Viral diseases represent a major public health concerns and ever-present risks for developing into future pandemics. Antiviral antibody therapeutics, either alone or in combination with other therapies, emerged as valuable preventative treatment options, including during global emergencies. Here we will discuss polyclonal monoclonal antiviral focusing on the unique biochemical physiological properties that make them well-suited therapeutic agents. We describe methods of characterization potency assessment throughout development, highlighting similarities differences between products appropriate. In addition, consider benefits challenges antibodies when used types therapeutics. Lastly, novel approaches to development identify areas would benefit from additional research.

Язык: Английский

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The S2 subunit of spike encodes diverse targets for functional antibody responses to SARS-CoV-2

PLoS Pathogens, Год журнала: 2024, Номер 20(8), С. e1012383 - e1012383

Опубликована: Авг. 2, 2024

The SARS-CoV-2 virus responsible for the COVID-19 global pandemic has exhibited a striking capacity viral evolution that drives continued evasion from vaccine and infection-induced immune responses. Mutations in receptor binding domain of S1 subunit spike glycoprotein have led to considerable escape antibody responses, reducing efficacy vaccines monoclonal (mAb) therapies. Therefore, there is need interrogate more constrained regions spike, such as S2 subdomain. Here, we present collection mAbs two convalescent individuals target multiple S2, including outside those commonly reported. One mAbs, C20.119, which bound highly conserved epitope fusion peptide, was able broadly neutralize across variants, SARS-CoV-1, closely related zoonotic sarbecoviruses. majority were non-neutralizing; however, many them could mediate antibody-dependent cellular cytotoxicity (ADCC) at levels similar S1-targeting mAb S309 previously authorized treatment infections. Several with ADCC function also trimers other human coronaviruses (HCoVs), MERS-CoV HCoV-HKU1. Our findings suggest can diverse epitopes functional HCoV sarbecovirus breadth likely functionally spike. These be developed potential future pandemics, while providing insight into ideal eliciting broad response.

Язык: Английский

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Antibodies that neutralize all current SARS-CoV-2 variants of concern by conformational locking

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2023, Номер unknown

Опубликована: Апрель 10, 2023

SUMMARY SARS-CoV-2 continues to evolve and evade most existing neutralizing antibodies, including all clinically authorized antibodies. We have isolated characterized two human monoclonal 12-16 12-19, which exhibited activities against variants tested, BQ.1.1 XBB.1.5. They also blocked infection in hamsters challenged with Omicron BA.1 intranasally. Structural analyses revealed both antibodies targeted a conserved quaternary epitope located at the interface between N-terminal domain subdomain 1, revealing previously unrecognized site of vulnerability on spike. These prevent viral receptor engagement by locking receptor-binding spike down conformation, novel mechanism virus neutralization for non-RBD Deep mutational scanning showed that could mutate escape but responsible mutations are rarely found circulating viruses. Antibodies 12-19 hold promise as prophylactic agents immunocompromised persons who do not respond robustly COVID-19 vaccines.

Язык: Английский

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