Identification of broad, potent antibodies to functionally constrained regions of SARS-CoV-2 spike following a breakthrough infection

Proceedings of the National Academy of Sciences, Journal Year: 2023, Volume and Issue: 120(23)

Published: May 30, 2023

The antiviral benefit of antibodies can be compromised by viral escape especially for rapidly evolving viruses. Therefore, durable, effective must both broad and potent to counter newly emerging, diverse strains. Discovery such is critically important SARS-CoV-2 as the global emergence new variants concern (VOC) has efficacy therapeutic vaccines. We describe a collection neutralizing monoclonal (mAbs) isolated from an individual who experienced breakthrough infection with Delta VOC. Four mAbs potently neutralize Wuhan-Hu-1 vaccine strain, VOC, also retain potency against Omicron VOCs through BA.4/BA.5 in pseudovirus-based authentic virus assays. Three recently circulating XBB.1.5 BQ.1.1 one neutralizes SARS-CoV-1. these was greater than all but that had been approved applications. target distinct epitopes on spike glycoprotein, three receptor-binding domain (RBD) invariant region downstream RBD subdomain 1 (SD1). pathways we defined at single amino acid resolution deep mutational scanning show they conserved, functionally constrained regions suggesting could incur fitness cost. Overall, are unique their breadth across VOCs, epitope specificity, include highly mAb targeting rare outside SD1.

Language: Английский

---

Evaluation of SARS-CoV-2 Antibody Response Between Paired Fingerprick (HemaPEN®) and Venepuncture Collected Samples in Children and Adults

Antibodies, Journal Year: 2025, Volume and Issue: 14(1), P. 13 - 13

Published: Feb. 5, 2025

Serological surveillance of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies is important to monitor population COVID-19 immunity. Dried blood spots (DBS) are a valuable method for serosurveys, particularly in remote settings and children. We compared the measurement SARS-CoV-2 spike-specific IgG paired samples collected using standard venepuncture (serum) hemaPEN® microsampling DBS device from children adults. A total 83 participants (10 months 65 years age), comprising COVID-positive -negative participants, were recruited. Paired serum assayed receptor-binding domain (RBD) Spike (S1) an established in-house ELISA. RBD S1 concentrations hemaPEN eluates non-parametric Wilcoxon matched-pairs signed ranked test. Pearson's correlation was used level agreement between assessed by Bland-Altman analysis. N = 41 adults (36 5 COVID-negative), 42 (37 COVID-positive, COVID-negative) have assayed. found moderate strong correlations (r 0.9472, p < 0.0001) proteins 0.6892, 0.0001). Similar results observed both adult paediatric populations. No significant differences S1-specific levels stored up 35 weeks at room temperature. Eluted showed high specificity sensitivity (100% 89.89%, respectively) with serum. The use sample collection feasible approach assessing serosurveillance studies,

Language: Английский

---

Development and Implementation of Dried Blood Spot-Based COVID-19 Serological Assays for Epidemiologic Studies

Microbiology Spectrum, Journal Year: 2022, Volume and Issue: 10(3)

Published: May 25, 2022

Serological surveillance studies of infectious diseases provide population-level estimates infection and antibody prevalence, generating crucial insight into immunity, risk factors leading to infection, effectiveness public health measures. These traditionally rely on detection pathogen-specific antibodies in samples derived from venipuncture, an expensive logistically challenging aspect serological surveillance. During the COVID-19 pandemic, guidelines implemented prevent spread SARS-CoV-2 made collection venous blood difficult at a time when serosurveillance was urgently needed. Dried spots (DBS) have generated interest as alternative for applications due their stability, low cost, ease collection; DBS can be self-generated via fingerprick by community members mailed ambient temperatures. Here, we detail development four DBS-based methods demonstrate implementation large survey 12 cities East Bay region San Francisco metropolitan area using at-home collection. We find that perform similarly plasma/serum enzyme-linked immunosorbent assays commercial assays. In addition, show reliably detect responses months postinfection track kinetics after vaccination. Implementation enabled valuable data our study population investigate changes seroprevalence over 8-month period. Our work makes strong argument studies, not just SARS-CoV-2, but any situation where phlebotomy is inaccessible. IMPORTANCE Estimation community-level or vaccination critical inform responses. Traditional invasive procedure amenable pandemic-related social-distancing are since they self-collected participants home do require refrigeration shipment storage. However, measure levels been widely utilized. comparable sampling method measured distinct The format longitudinal cohort samples, ensuring participants' safety during ongoing pandemic. demonstrates excellent measuring whenever venipuncture impractical.

Language: Английский

---

Comparative performance data for multiplex SARS-CoV-2 serological assays from a large panel of dried blood spot specimens

Heliyon, Journal Year: 2022, Volume and Issue: 8(9), P. e10270 - e10270

Published: Aug. 28, 2022

The extent of the COVID-19 pandemic will be better understood through serosurveys and SARS-CoV-2 antibody testing. Dried blood spot (DBS) samples play a central role in large scale serosurveillance by simplifying biological specimen collection transportation, especially Canada. Direct comparative performance data on multiplex assays resulting from identical DBS are currently lacking. In our study, we aimed to provide for BioPlex 2200 IgG (Bio-Rad), V-PLEX Panel 2 (MSD), Elecsys Anti-SARS-CoV-2 (Roche) commercial assays, as well two highly scalable in-house (University Ottawa Mount Sinai Hospital protocols) assess their suitability DBS-based serosurveillance. These were evaluated against panels collected convalescent patients (

Language: Английский

---

Quantifying neutralising antibody responses against SARS-CoV-2 in dried blood spots (DBS) and paired sera

Scientific Reports, Journal Year: 2023, Volume and Issue: 13(1)

Published: Sept. 11, 2023

Abstract The ongoing SARS-CoV-2 pandemic was initially managed by non-pharmaceutical interventions such as diagnostic testing, isolation of positive cases, physical distancing and lockdowns. advent vaccines has provided crucial protection against SARS-CoV-2. Neutralising antibody (nAb) responses are a key correlate protection, therefore measuring nAb is essential for monitoring vaccine efficacy. Fingerstick dried blood spots (DBS) ideal use in large-scale sero-surveillance because they inexpensive, offer the option self-collection can be transported stored at ambient temperatures. Such advantages also make DBS appealing to resource-limited settings potential future pandemics. In this study, sera, venous fingerstick on filter paper were measured. Samples collected from acutely infected individuals, convalescent individuals vaccinated individuals. Good agreement observed between measured eluted paired sera. Stability sera room temperature 28 days. Overall, study provides support viable sample collection method responses.

Language: Английский

---

A bead-based multiplex assay covering all coronaviruses pathogenic for humans for sensitive and specific surveillance of SARS-CoV-2 humoral immunity

Scientific Reports, Journal Year: 2023, Volume and Issue: 13(1)

Published: Dec. 9, 2023

Serological assays measuring antibodies against SARS-CoV-2 are key to describe the epidemiology, pathobiology or induction of immunity after infection vaccination. Of those, multiplex targeting multiple antigens especially helpful as closely related coronaviruses other can be analysed simultaneously from small sample volumes, hereby shedding light on patterns in immune response that would otherwise remain undetected. We established a bead-based 17-plex assay detecting all pathogenic for humans: SARS-CoV-2, SARS-CoV, MERS-CoV, HCoV strains 229E, OC43, HKU1, and NL63. The was validated five commercial serological immunoassays, surrogate virus neutralisation test, assay, SARS-CoV-2. It found highly versatile shown by antibody detection both serum dried blot spots three case studies. First, we followed seroconversion four endemic an outbreak study day-care centres children. Second, were able link more severe clinical course stronger IgG with this 17-plex-assay, which IgG1 IgG3 dominated. Finally, our discriminate recent previous infections calculating IgG/IgM ratio N antigen antibodies. In conclusion, due comprehensive method comparison, thorough validation, proven versatility, is valuable tool studies coronavirus serology.

Language: Английский

---

Applications of SARS-CoV-2 serological testing: impact of test performance, sample matrices, and patient characteristics

Critical Reviews in Clinical Laboratory Sciences, Journal Year: 2023, Volume and Issue: 61(1), P. 70 - 88

Published: Oct. 6, 2023

AbstractLaboratory testing has been a key tool in managing the SARS-CoV-2 global pandemic. While rapid antigen and PCR proven useful for diagnosing acute infections, additional methods are required to understand long-term impact of infections on immune response. Serological testing, well-documented laboratory practice, measures presence antibodies sample uncover information about host immunity. Although proposed applications serological clinical use have previously limited, current research into shown growing utility these settings. To name few, used identify patients with past active disease monitor vaccine efficacy. Test result interpretation, however, often complicated by factors that include poor test sensitivity early infection, lack response some individuals, overlying infection vaccination responses, standardization antibody titers/levels between instruments, unknown titers confer protection, large between-individual biological variation following or vaccination. Thus, three major components this review will examine (1) affect utility: performance, matrices, seroprevalence concerns viral variants, (2) patient response: timing sampling, age, sex, body mass index, immunosuppression vaccination, (3) informative testing: identifying surveillance guide health practices, examination protective should be beneficial care if it is implemented appropriately. However, as other developed tests, serology modality warrants careful consideration limitations evaluation its utility.Keywords: SARS-CoV-2COVID-19serologyimmunityantibody Disclosure statementJT receives in-kind support from Roche Diagnostics.Additional informationFundingThis open-access article supported funding Canadian Institutes Health Research (Funding Reference Number VR4-172753, VS1-175526, VS2-175572.

Language: Английский

---

RSV Neutralizing Antibodies in Dried Blood

The Journal of Infectious Diseases, Journal Year: 2023, Volume and Issue: 230(1), P. e93 - e101

Published: Nov. 28, 2023

The key correlate of protection respiratory syncytial virus (RSV) vaccines and monoclonal antibodies (mAb) is neutralization, measured using sera obtained through venipuncture. Dried blood with a finger prick can simplify acquisition, processing, storage, transport in trials, thereby reduce costs. In this study we validate an assay to measure RSV neutralization dried capillary blood.

Language: Английский

---

Identification of broad, potent antibodies to functionally constrained regions of SARS-CoV-2 spike following a breakthrough infection

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2022, Volume and Issue: unknown

Published: Dec. 16, 2022

The antiviral benefit of antibodies can be compromised by viral escape especially for rapidly evolving viruses. Therefore, durable, effective must both broad and potent to counter newly emerging, diverse strains. Discovery such is critically important SARS-CoV-2 as the global emergence new variants concern (VOC) has efficacy therapeutic vaccines. We describe a collection neutralizing monoclonal (mAbs) isolated from an individual who experienced breakthrough infection with Delta VOC. Four mAbs potently neutralize Wuhan-Hu-1 vaccine strain, VOC, also retain potency against Omicron VOCs through BA.4/BA.5 in pseudovirus-based authentic virus assays. Three recently circulating XBB.1.5 BQ.1.1 one neutralizes SARS-CoV-1. these was greater than all but that had been approved applications. target distinct epitopes on spike glycoprotein, three receptor binding domain (RBD) invariant region downstream RBD subdomain 1 (SD1). pathways we defined at single amino acid resolution deep mutational scanning show they conserved, functionally constrained regions suggesting could incur fitness cost. Overall, are novel their breadth across VOCs, epitope specificity, include highly mAb targeting rare outside SD1.

Language: Английский

---

Low pre-infection levels of neutralizing antibody in breakthrough infections after bivalent BA.4–5 vaccine and practical application of dried blood spots

Vaccine, Journal Year: 2024, Volume and Issue: 42(22), P. 126029 - 126029

Published: June 4, 2024

The level of neutralizing antibodies required to confer protection against COVID-19 breakthrough infections (BIs) is unclear, and the ability know immune status individuals rapidly changing endemic variants limited. We assessed longitudinal serum anti-RBD antibody levels activities (NTs) Omicron BA.5 XBB.1.5 in healthcare workers following fourth monovalent fifth bivalent BA.4-5 vaccines. occurrence BIs was also followed, pre-infection were compared between patients who developed BI those did not. In addition, we collected whole blood samples on same day as sera stored them filter papers (nos. 545, 590, 424) for up two months, then measured their NTs using dried spots (DBS) eluates, with paired sera. Pre-infection lower than not, but not different them. below 50 % 200-fold diluted might be one indicators high risk BI. However, at 6 months after dose vaccine this threshold almost half infection-naïve participants. DBS eluates strongly correlated sera, time temperature stability varied type paper; no. 545 paper found most suitable NT evaluation.

Language: Английский

---