scholarly journals Molecular clamp stabilised Spike protein for protection against SARS-CoV-2

2020 ◽  
Author(s):  
Daniel Watterson ◽  
Danushka Wijesundara ◽  
Naphak Modhiran ◽  
Francesca Mordant ◽  
Zheyi Li ◽  
...  
Keyword(s):  
Clinical Evaluation ◽  
Large Scale ◽  
Vaccine Candidate ◽  
Receptor Binding Domain ◽  
Preclinical Development ◽  
Neutralising Antibodies ◽  
Clinical Safety ◽  
Toxicology Study ◽  
Efficacy Trials ◽  
Large Scale Manufacture

Abstract Efforts to develop and deploy effective vaccines against Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) continue at pace with more than 30 candidate vaccines now in clinical evaluation. Here we describe the preclinical development of an adjuvanted, prefusion-stabilised Spike (S) protein “Sclamp” subunit vaccine, from rational antigen design through to assessing manufacturability and vaccine efficacy. In mice, the vaccine candidate elicits high levels of neutralising antibodies to epitopes both within and outside the receptor binding domain (RBD) of S, as well as broadly reactive and polyfunctional S-specific CD4+ and cytotoxic CD8+ T cells. We also show protection in Syrian hamsters, which has emerged as a robust animal model for pulmonary SARS-CoV-2 infection. No evidence of vaccine enhanced disease was observed in animal challenge studies and pre-clinical safety was further demonstrated in a GLP toxicology study in rats. The Sclamp vaccine candidate is currently progressing rapidly through clinical evaluation in parallel with large-scale manufacture for pivotal efficacy trials and potential widespread distribution.

scholarly journals Neutralising SARS-CoV-2 RBD-specific antibodies persist for at least six months independently of symptoms in adults

2021 ◽  
Vol 1 (1) ◽  
Author(s):  
Angelika Wagner ◽  
Angela Guzek ◽  
Johanna Ruff ◽  
Joanna Jasinska ◽  
Ute Scheikl ◽  
...  
Keyword(s):  
Nucleocapsid Protein ◽  
Large Scale ◽  
Kappa Coefficient ◽  
Receptor Binding Domain ◽  
Neutralising Antibodies ◽  
Specific Antibodies ◽  
Time Points ◽  
Large Scale Testing ◽  
Iga Antibodies ◽  
Set Up

Abstract Background In spring 2020, at the beginning of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic in Europe, we set up an assay system for large-scale testing of virus-specific and neutralising antibodies including their longevity. Methods We analysed the sera of 1655 adult employees for SARS-CoV-2-specific antibodies using the S1 subunit of the spike protein of SARS-CoV-2. Sera containing S1-reactive antibodies were further evaluated for receptor-binding domain (RBD)- and nucleocapsid protein (NCP)-specific antibodies in relation to the neutralisation test (NT) results at three time points over six months. Results We detect immunoglobulin G (IgG) and/or IgA antibodies reactive to the S1 protein in 10.15% (n = 168) of the participants. In total, 0.97% (n = 16) are positive for S1-IgG, 0.91% (n = 15) were S1-IgG- borderline and 8.28% (n = 137) exhibit only S1-IgA antibodies. Of the 168 S1-reactive sera, 8.33% (n = 14) have detectable RBD-specific antibodies and 6.55% (n = 11) NCP-specific antibodies. The latter correlates with NTs (kappa coefficient = 0.8660) but start to decline after 3 months. RBD-specific antibodies correlate most closely with the NT (kappa = 0.9448) and only these antibodies are stable for up to six months. All participants with virus-neutralising antibodies report symptoms, of which anosmia and/or dysgeusia correlate most closely with the detection of virus-neutralising antibodies. Conclusions RBD-specific antibodies are most reliably detected post-infection, independent of the number/severity of symptoms, and correlate with neutralising antibodies at least for six months. They thus qualify best for large-scale seroepidemiological evaluation of both antibody reactivity and virus neutralisation.

scholarly journals A tandem-repeat dimeric RBD protein-based COVID-19 vaccine ZF2001 protects mice and nonhuman primates

2021 ◽  
Author(s):  
Yaling An ◽  
Shihua Li ◽  
Xiyue Jin ◽  
Jian-bao Han ◽  
Kun Xu ◽  
...  
Keyword(s):  
Tandem Repeat ◽  
Nonhuman Primates ◽  
Vaccine Candidate ◽  
Neutralizing Antibody ◽  
Cellular Responses ◽  
Receptor Binding Domain ◽  
Preclinical Development ◽  
Phase 3 ◽  
Lung Injuries ◽  
Trachea And Bronchi

AbstractA safe, efficacious and deployable vaccine is urgently needed to control COVID-19 pandemic. We report here the preclinical development of a COVID-19 vaccine candidate, ZF2001, which contains tandem-repeat dimeric receptor-binding domain (RBD) protein with alum-based adjuvant. We assessed vaccine immunogenicity and efficacy in both mice and non-human primates (NHPs). ZF2001 induced high levels of RBD-binding and SARS-CoV-2 neutralizing antibody in both mice and NHPs, and also elicited balanced TH1/TH2 cellular responses in NHPs. Two doses of ZF2001 protected Ad-hACE2-transduced mice against SARS-CoV-2 infection, as detected by reduced viral RNA and relieved lung injuries. In NHPs, vaccination of either 25 μg or 50 μg ZF2001 prevented infection with SARS-CoV-2 in lung, trachea and bronchi, with milder lung lesions. No evidence of disease enhancement is observed in both models. ZF2001 is being evaluated in the ongoing international multi-center Phase 3 trials (NCT04646590) and has been approved for emergency use in Uzbekistan.

scholarly journals Safety and immunogenicity of an mRNA-lipid nanoparticle vaccine candidate against SARS-CoV-2

2021 ◽  
Author(s):  
Peter G. Kremsner ◽  
Philipp Mann ◽  
Arne Kroidl ◽  
Isabel Leroux-Roels ◽  
Christoph Schindler ◽  
...  
Keyword(s):  
Immune Responses ◽  
Vaccine Candidate ◽  
Phase 1 ◽  
Lipid Nanoparticles ◽  
Enzyme Linked Immunosorbent Assay ◽  
Receptor Binding Domain ◽  
Igg Antibodies ◽  
Phase 1 Study ◽  
S Protein ◽  
Dosage Escalation

Summary Background We used the RNActive® technology platform (CureVac N.V., Tübingen, Germany) to prepare CVnCoV, a COVID-19 vaccine containing sequence-optimized mRNA coding for a stabilized form of SARS-CoV‑2 spike (S) protein encapsulated in lipid nanoparticles (LNP). Methods This is an interim analysis of a dosage escalation phase 1 study in healthy 18–60-year-old volunteers in Hannover, Munich and Tübingen, Germany, and Ghent, Belgium. After giving 2 intramuscular doses of CVnCoV or placebo 28 days apart we assessed solicited local and systemic adverse events (AE) for 7 days and unsolicited AEs for 28 days after each vaccination. Immunogenicity was measured as enzyme-linked immunosorbent assay (ELISA) IgG antibodies to SARS-CoV‑2 S‑protein and receptor binding domain (RBD), and SARS-CoV‑2 neutralizing titers (MN50). Results In 245 volunteers who received 2 CVnCoV vaccinations (2 μg, n = 47, 4 μg, n = 48, 6 μg, n = 46, 8 μg, n = 44, 12 μg, n = 28) or placebo (n = 32) there were no vaccine-related serious AEs. Dosage-dependent increases in frequency and severity of solicited systemic AEs, and to a lesser extent local AEs, were mainly mild or moderate and transient in duration. Dosage-dependent increases in IgG antibodies to S‑protein and RBD and MN50 were evident in all groups 2 weeks after the second dose when 100% (23/23) seroconverted to S‑protein or RBD, and 83% (19/23) seroconverted for MN50 in the 12 μg group. Responses to 12 μg were comparable to those observed in convalescent sera from known COVID-19 patients. Conclusion In this study 2 CVnCoV doses were safe, with acceptable reactogenicity and 12 μg dosages elicited levels of immune responses that overlapped those observed in convalescent sera.

scholarly journals Enhanced Neutralizing Antibody Response Induced by Respiratory Syncytial Virus Prefusion F Protein Expressed by a Vaccine Candidate

2015 ◽  
Vol 89 (18) ◽  
pp. 9499-9510 ◽  
Author(s):  
Bo Liang ◽  
Sonja Surman ◽  
Emerito Amaro-Carambot ◽  
Barbora Kabatova ◽  
Natalie Mackow ◽  
...  
Keyword(s):  
Respiratory Syncytial Virus ◽  
Clinical Evaluation ◽  
Genetic Stability ◽  
Vaccine Candidate ◽  
Early Passage ◽  
F Protein ◽  
Syncytial Virus ◽  
Human Parainfluenza Virus ◽  
Type 3

ABSTRACTRespiratory syncytial virus (RSV) and human parainfluenza virus type 3 (HPIV3) are the first and second leading viral agents of severe respiratory tract disease in infants and young children worldwide. Vaccines are not available, and an RSV vaccine is particularly needed. A live attenuated chimeric recombinant bovine/human PIV3 (rB/HPIV3) vector expressing the RSV fusion (F) glycoprotein from an added gene has been under development as a bivalent vaccine against RSV and HPIV3. Previous clinical evaluation of this vaccine candidate suggested that increased genetic stability and immunogenicity of the RSV F insert were needed. This was investigated in the present study. RSV F expression was enhanced 5-fold by codon optimization and by modifying the amino acid sequence to be identical to that of an early passage of the original clinical isolate. This conferred a hypofusogenic phenotype that presumably reflects the original isolate. We then compared vectors expressing stabilized prefusion and postfusion versions of RSV F. In a hamster model, prefusion F induced increased quantity and quality of RSV-neutralizing serum antibodies and increased protection against wild-type (wt) RSV challenge. In contrast, a vector expressing the postfusion F was less immunogenic and protective. The genetic stability of the RSV F insert was high and was not affected by enhanced expression or the prefusion or postfusion conformation of RSV F. These studies provide an improved version of the previously well-tolerated rB/HPIV3-RSV F vaccine candidate that induces a superior RSV-neutralizing serum antibody response.IMPORTANCERespiratory syncytial virus (RSV) and human parainfluenza virus type 3 (HPIV3) are two major causes of pediatric pneumonia and bronchiolitis. The rB/HPIV3 vector expressing RSV F protein is a candidate bivalent live vaccine against HPIV3 and RSV. Previous clinical evaluation indicated the need to increase the immunogenicity and genetic stability of the RSV F insert. Here, we increased RSV F expression by codon optimization and by modifying the RSV F amino acid sequence to conform to that of an early passage of the original isolate. This resulted in a hypofusogenic phenotype, which likely represents the original phenotype before adaptation to cell culture. We also included stabilized versions of prefusion and postfusion RSV F protein. Prefusion RSV F induced a larger quantity and higher quality of RSV-neutralizing serum antibodies and was highly protective. This provides an improved candidate for further clinical evaluation.

scholarly journals Metachronal waves in magnetic micro-robotic paddles for artificial cilia

2021 ◽  
Vol 2 (1) ◽  
Author(s):  
Matthew T. Bryan ◽  
Elizabeth L. Martin ◽  
Aleksandra Pac ◽  
Andrew D. Gilbert ◽  
Feodor Y. Ogrin
Keyword(s):  
Large Scale ◽  
Point Of Care ◽  
Hydrodynamic Response ◽  
Flow Speeds ◽  
Break Time ◽  
Large Scale Manufacture ◽  
Fresh Insight ◽  
Metachronal Waves ◽  
Insight Into ◽  
Phase Differences

AbstractBiological cilia generate fluid movement within viscosity-dominated environments using beating motions that break time-reversal symmetry. This creates a metachronal wave, which enhances flow efficiency. Artificially mimicking this behaviour could improve microfluidic point-of-care devices, since viscosity-dominated fluid dynamics impede fluid flow and mixing of reagents, limiting potential for multiplexing diagnostic tests. However, current biomimicry schemes require either variation in the hydrodynamic response across a cilia array or a complex magnetic anisotropy configuration to synchronise the actuation sequence with the driving field. Here, we show that simple modifications to the structural design introduce phase differences between individual actuators, leading to the spontaneous formation of metachronal waves. This generates flow speeds of up to 16 μm/s as far as 675 μm above the actuator plane. By introducing metachronal waves through lithographic structuring, large scale manufacture becomes feasible. Additionally, by demonstrating that metachronal waves emerge from non-uniformity in internal structural mechanics, we offer fresh insight into the mechanics of cilia coordination.

scholarly journals Towards large-scale identification of HLA-restricted T cell epitopes from four vaccine candidate antigens in a malaria endemic community in Ghana

Vaccine ◽  
2021 ◽  
Author(s):  
Kwadwo Asamoah Kusi ◽  
Ebenezer Addo Ofori ◽  
Kwadwo Akyea-Mensah ◽  
Eric Kyei-Baafour ◽  
Augustina Frimpong ◽  
...  
Keyword(s):  
T Cell ◽  
Large Scale ◽  
Vaccine Candidate ◽  
T Cell Epitopes

scholarly journals SARS-CoV-2 B.1.617 emergence and sensitivity to vaccine-elicited antibodies

2021 ◽  
Author(s):  
Isabella Ferreira ◽  
Rawlings Datir ◽  
Guido Papa ◽  
Steven Kemp ◽  
Bo Meng ◽  
...  
Keyword(s):  
Receptor Binding ◽  
Growth Rates ◽  
Cleavage Site ◽  
Syncytium Formation ◽  
Binding Domain ◽  
Spike Protein ◽  
Receptor Binding Domain ◽  
Additive Effects ◽  
Neutralising Antibodies ◽  
Indian State

The B.1.617 variant emerged in the Indian state of Maharashtra in late 2020 and has spread throughout India and to at least 40 countries. There have been fears that two key mutations seen in the receptor binding domain L452R and E484Q would have additive effects on evasion of neutralising antibodies. Here we delineate the phylogenetics of B.1.617 and spike mutation frequencies, in the context of others bearing L452R. The defining mutations in B.1.617.1 spike are L452R and E484Q in the RBD that interacts with ACE2 and is the target of neutralising antibodies. All B.1.617 viruses have the P681R mutation in the polybasic cleavage site region in spike. We report that B.1.617.1 spike bearing L452R, E484Q and P681R mediates entry into cells with slightly reduced efficiency compared to Wuhan-1. This spike confers modestly reduced sensitivity to BNT162b2 mRNA vaccine-elicited antibodies that is similar in magnitude to the loss of sensitivity conferred by L452R or E484Q alone. Furthermore we show that the P681R mutation significantly augments syncytium formation upon the B.1.617.1 spike protein, potentially contributing to increased pathogenesis observed in hamsters and infection growth rates observed in humans.

scholarly journals Adaptation of SARS-CoV-2 in BALB/c mice for testing vaccine efficacy

Science ◽  
2020 ◽  
pp. eabc4730 ◽  
Author(s):  
Hongjing Gu ◽  
Qi Chen ◽  
Guan Yang ◽  
Lei He ◽  
Hang Fan ◽  
...  
Keyword(s):  
Vaccine Candidate ◽  
Inflammatory Responses ◽  
Protective Efficacy ◽  
Small Animal ◽  
Spike Protein ◽  
Mouse Lung ◽  
Receptor Binding Domain ◽  
Intranasal Inoculation ◽  
Adaptive Mutations ◽  
Small Animal Models

The ongoing COVID-19 pandemic has prioritized the development of small animal models for SARS-CoV-2. Herein, we adapted a clinical isolate of SARS-CoV-2 by serial passaging in the respiratory tract of aged BALB/c mice. The resulting mouse-adapted strain at passage 6 (termed MASCp6) showed increased infectivity in mouse lung, and led to interstitial pneumonia and inflammatory responses in both young and aged mice following intranasal inoculation. Deep sequencing revealed a panel of adaptive mutations potentially associated with the increased virulence. In particular, the N501Y mutation is located at the receptor binding domain (RBD) of the spike protein. The protective efficacy of a recombinant RBD vaccine candidate was validated using this model. Thus, this mouse-adapted strain and associated challenge model should be of value in evaluating vaccines and antivirals against SARS-CoV-2.

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