Visual and High-Efficiency Secretion of SARS-CoV-2 Nanobodies with Escherichia coli DOI Creative Commons
Shuai Zhao,

Wanting Zeng,

Fang Yu

et al.

Biomolecules, Journal Year: 2025, Volume and Issue: 15(1), P. 111 - 111

Published: Jan. 12, 2025

Nanobodies have gained attention as potential therapeutic and diagnostic agents for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) due to their ability bind neutralize the virus. However, rapid, scalable, robust production of nanobodies SARS-CoV-2 remains a crucial challenge. In this study, we developed visual high-efficiency biomanufacturing method with Escherichia coli by fusing super-folder green fluorescent protein (sfGFP) N-terminus or C-terminus nanobody. Several receptor-binding domain (RBD)-specific spike (S) were secreted onto surface E. cells even into culture medium, including Fu2, ANTE, mNb6, MR3-MR3, n3113.1. The retained equal activity prior research, regardless whether sfGFP was removed. Since some bound different regions RBD, combined two improve affinity. Fu2-sfGFP-ANTE constructed be bispecific nanobody exhibited significantly higher affinity than Fu2 (35.0-fold), ANTE (7.3-fold), combination (3.3-fold). Notably, can normally medium outer membrane. novel system enhances efficiency expression streamlines downstream purification process, enabling large-scale, cost-effective production. addition, secreting on facilitates screening characterization antigen-binding clones.

Language: Английский

Antibody escape of SARS-CoV-2 Omicron BA.4 and BA.5 from vaccine and BA.1 serum DOI Creative Commons
Aekkachai Tuekprakhon, Rungtiwa Nutalai, Aiste Dijokaite-Guraliuc

et al.

Cell, Journal Year: 2022, Volume and Issue: 185(14), P. 2422 - 2433.e13

Published: June 9, 2022

The Omicron lineage of SARS-CoV-2, which was first described in November 2021, spread rapidly to become globally dominant and has split into a number sublineages. BA.1 dominated the initial wave but been replaced by BA.2 many countries. Recent sequencing from South Africa's Gauteng region uncovered two new sublineages, BA.4 BA.5, are taking over locally, driving wave. BA.5 contain identical spike sequences, although closely related BA.2, they further mutations receptor-binding domain their spikes. Here, we study neutralization BA.4/5 using range vaccine naturally immune serum panels monoclonal antibodies. shows reduced individuals vaccinated with triple doses AstraZeneca or Pfizer compared BA.2. Furthermore, breakthrough infections, there are, likewise, significant reductions BA.4/5, raising possibility repeat infections.

Language: Английский

Citations

676
Structural biology of SARS-CoV-2 and implications for therapeutic development DOI Creative Commons
Haitao Yang, Zihe Rao

Nature Reviews Microbiology, Journal Year: 2021, Volume and Issue: 19(11), P. 685 - 700

Published: Sept. 17, 2021

Language: Английский

Citations

416
Versatile and multivalent nanobodies efficiently neutralize SARS-CoV-2 DOI Creative Commons
Yufei Xiang, Sham Nambulli, Zhengyun Xiao

et al.

Science, Journal Year: 2020, Volume and Issue: 370(6523), P. 1479 - 1484

Published: Nov. 6, 2020

Nanobodies that neutralize Monoclonal antibodies bind to the spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) show therapeutic promise but must be produced in mammalian cells and need delivered intravenously. By contrast, single-domain called nanobodies can bacteria or yeast, their stability may enable aerosol delivery. Two papers now report tightly efficiently SARS-CoV-2 cells. Schoof et al. screened a yeast surface display synthetic Xiang anti-spike by llama. Both groups identified highly potent lock an inactive conformation. Multivalent constructs selected achieved even more neutralization. Science , this issue p. 1473 1479

Language: Английский

Citations

407
LY-CoV1404 (bebtelovimab) potently neutralizes SARS-CoV-2 variants DOI Creative Commons
Kathryn Westendorf,

Stefanie Žentelis,

Lingshu Wang

et al.

Cell Reports, Journal Year: 2022, Volume and Issue: 39(7), P. 110812 - 110812

Published: April 25, 2022

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-neutralizing monoclonal antibodies (mAbs) can reduce the risk of hospitalization from disease 2019 (COVID-19) when administered early. However, SARS-CoV-2 variants concern (VOCs) have negatively affected therapeutic use some authorized mAbs. Using a high-throughput B cell screening pipeline, we isolated LY-CoV1404 (bebtelovimab), highly potent spike glycoprotein receptor binding domain (RBD)-specific antibody. potently neutralizes authentic SARS-CoV-2, B.1.1.7, B.1.351, and B.1.617.2. In pseudovirus neutralization studies, variants, including B.1.617.2, B.1.427/B.1.429, P.1, B.1.526, B.1.1.529, BA.2 subvariant. Structural analysis reveals that contact residues epitope are conserved, except for N439 N501. The neutralizing activity is unaffected by most common mutations at these positions (N439K N501Y). broad relatively conserved suggest has potential to be an effective agent treat all known variants.

Language: Английский

Citations

402
Role of Structural and Non-Structural Proteins and Therapeutic Targets of SARS-CoV-2 for COVID-19 DOI Creative Commons
Rohitash Yadav, Jitendra Kumar Chaudhary, Neeraj Jain

et al.

Cells, Journal Year: 2021, Volume and Issue: 10(4), P. 821 - 821

Published: April 6, 2021

Coronavirus belongs to the family of Coronaviridae, comprising single-stranded, positive-sense RNA genome (+ ssRNA) around 26 32 kilobases, and has been known cause infection a myriad mammalian hosts, such as humans, cats, bats, civets, dogs, camels with varied consequences in terms death debilitation. Strikingly, novel coronavirus (2019-nCoV), later renamed severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), found be causative agent disease-19 (COVID-19), shows 88% sequence identity bat-SL-CoVZC45 bat-SL-CoVZXC21, 79% SARS-CoV 50% MERS-CoV, respectively. Despite key amino acid residual variability, there is an incredible structural similarity between receptor binding domain (RBD) spike protein (S) SARS-CoV-2 SARS-CoV. During infection, compared displays 10–20 times greater affinity for its cognate host cell receptor, angiotensin-converting enzyme 2 (ACE2), leading proteolytic cleavage S by transmembrane protease serine (TMPRSS2). Following cellular entry, ORF-1a ORF-1ab, located downstream 5′ end + ssRNA genome, undergo translation, thereby forming two large polyproteins, pp1a pp1ab. These following protease-induced molecular assembly, form functional viral polymerase, also referred replicase. Thereafter, uninterrupted orchestrated replication-transcription events lead synthesis multiple nested sets subgenomic mRNAs (sgRNAs), which are finally translated several accessory proteins participating structure formation various functions virus, assemble encapsulate genomic (gRNA), resulting numerous progenies, eventually exit cell, spread rest body. In this review, we primarily focus on organization, non-structural components, potential prospective targets development therapeutic drugs, convalescent plasm therapy, vaccines tackle infection.

Language: Английский

Citations

385
Structure-guided multivalent nanobodies block SARS-CoV-2 infection and suppress mutational escape DOI Creative Commons
Paul-Albert Koenig, Hrishikesh Das,

Hejun Liu

et al.

Science, Journal Year: 2021, Volume and Issue: 371(6530)

Published: Jan. 12, 2021

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to spread, with devastating consequences. For passive immunization efforts, nanobodies have size and cost advantages over conventional antibodies. In this study, we generated four neutralizing that target the receptor binding domain of SARS-CoV-2 spike protein. We used x-ray crystallography cryo-electron microscopy define two distinct epitopes. On basis these structures, engineered multivalent more than 100 times activity monovalent nanobodies. Biparatopic nanobody fusions suppressed emergence escape mutants. Several constructs neutralized through competition, whereas other biparatopic triggered aberrant activation fusion machinery. These premature conformational changes in protein forestalled productive rendered virions noninfectious.

Language: Английский

Citations

371
An alpaca nanobody neutralizes SARS-CoV-2 by blocking receptor interaction DOI Creative Commons
Leo Hanke, Laura Perez Vidakovics, Daniel J. Sheward

et al.

Nature Communications, Journal Year: 2020, Volume and Issue: 11(1)

Published: Sept. 4, 2020

Abstract SARS-CoV-2 enters host cells through an interaction between the spike glycoprotein and angiotensin converting enzyme 2 (ACE2) receptor. Directly preventing this presents attractive possibility for suppressing replication. Here, we report isolation characterization of alpaca-derived single domain antibody fragment, Ty1, that specifically targets receptor binding (RBD) spike, directly ACE2 engagement. Ty1 binds RBD with high affinity, occluding ACE2. A cryo-electron microscopy structure bound complex at 2.9 Å resolution reveals to epitope on accessible in both ‘up’ ‘down’ conformations, sterically hindering RBD-ACE2 binding. While fusion Fc renders extremely potent, neutralizes pseudovirus as a 12.8 kDa nanobody, which can be expressed quantities bacteria, presenting opportunities manufacturing scale. is therefore excellent candidate intervention against COVID-19.

Language: Английский

Citations

324
Structural basis for the neutralization of SARS-CoV-2 by an antibody from a convalescent patient DOI Creative Commons
Daming Zhou, Helen M. E. Duyvesteyn, Cheng‐Pin Chen

et al.

Nature Structural & Molecular Biology, Journal Year: 2020, Volume and Issue: 27(10), P. 950 - 958

Published: July 31, 2020

Language: Английский

Citations

313
Molecular mechanism of interaction between SARS-CoV-2 and host cells and interventional therapy DOI Creative Commons
Qianqian Zhang, Rong Xiang, Shanshan Huo

et al.

Signal Transduction and Targeted Therapy, Journal Year: 2021, Volume and Issue: 6(1)

Published: June 11, 2021

Abstract The pandemic of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome 2 (SARS-CoV-2) infection has resulted in an unprecedented setback for global economy and health. SARS-CoV-2 exceptionally high level transmissibility extremely broad tissue tropism. However, the underlying molecular mechanism responsible sustaining this degree virulence remains largely unexplored. In article, we review current knowledge crucial information about how attaches on surface host cells through a variety receptors, such as ACE2, neuropilin-1, AXL, antibody–FcγR complexes. We further explain its spike (S) protein undergoes conformational transition from prefusion to postfusion with help proteases like furin, TMPRSS2, cathepsins. then ongoing experimental studies clinical trials antibodies, peptides, or small-molecule compounds anti-SARS-CoV-2 activity, discuss these antiviral therapies targeting host–pathogen interaction could potentially suppress viral attachment, reduce exposure fusion peptide curtail membrane block formation six-helix bundle (6-HB) core. Finally, specter rapidly emerging variants deserves serious broad-spectrum drugs vaccines long-term prevention control COVID-19 future.

Language: Английский

Citations

308
In vitro and in vivo functions of SARS-CoV-2 infection-enhancing and neutralizing antibodies DOI Creative Commons
Dapeng Li, Robert J. Edwards, Kartik Manne

et al.

Cell, Journal Year: 2021, Volume and Issue: 184(16), P. 4203 - 4219.e32

Published: June 18, 2021

Language: Английский

Citations

281