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Article ; Online: Developing antibody tests for SARS-CoV-2.

Petherick, Anna

2020 Volume 395, Issue 10230, Page(s) 1101–1102

MeSH term(s)	Antibodies, Viral/metabolism ; Betacoronavirus/immunology ; Betacoronavirus/isolation & purification ; COVID-19 ; Coronavirus Infections/diagnosis ; Humans ; Pandemics ; Pneumonia, Viral/diagnosis ; SARS-CoV-2
Chemical Substances	Antibodies, Viral
Keywords	covid19
Language	English
Publishing date	2020-04-02
Publishing country	England
Document type	Journal Article
ZDB-ID	3306-6
ISSN	1474-547X ; 0023-7507 ; 0140-6736
ISSN (online)	1474-547X
ISSN	0023-7507 ; 0140-6736
DOI	10.1016/S0140-6736(20)30788-1
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Article: Developing antibody tests for SARS-CoV-2

Petherick, Anna

Lancet

Keywords	covid19
Publisher	WHO
Document type	Article
Note	WHO #Covidence: #31283
Database	COVID19

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Article ; Online: Developing antibody tests for SARS-CoV-2

Petherick, Anna

The Lancet

2020 Volume 395, Issue 10230, Page(s) 1101–1102

Keywords	General Medicine ; covid19
Language	English
Publisher	Elsevier BV
Publishing country	us
Document type	Article ; Online
ZDB-ID	3306-6
ISSN	1474-547X ; 0023-7507 ; 0140-6736
ISSN (online)	1474-547X
ISSN	0023-7507 ; 0140-6736
DOI	10.1016/s0140-6736(20)30788-1
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Article: SARS-CoV-2 Rapid Antigen Test Based on a New Anti-Nucleocapsid Protein Monoclonal Antibody: Development and Real-Time Validation.

Coelho, Fabiana Fioravante / da Silva, Miriam Aparecida / Lopes, Thiciany Blener / Polatto, Juliana Moutinho / de Castro, Natália Salazar / Andrade, Luis Adan Flores / Lourenço, Karine Lima / Sato, Hugo Itaru / de Carvalho, Alex Fiorini / Coelho, Helena Perez / Bagno, Flávia Fonseca / Luz, Daniela / Viala, Vincent Louis / Cattony, Pedro Queiroz / Melo, Bruna de Sousa / Moro, Ana Maria / Quintilio, Wagner / Barbosa, Ana Paula / Bomfim, Camila Gasque /

Soares, Camila Pereira / Guzzo, Cristiane Rodrigues / Fonseca, Flavio Guimarães / Durigon, Edison Luiz / Gazzinelli, Ricardo Tostes / Ribeiro Teixeira, Santuza M / Piazza, Roxane Maria Fontes / Fernandes, Ana Paula

Microorganisms

2023 Volume 11, Issue 10

Abstract: ... used. However, as SARS-CoV-2 variants may continuously emerge, the replacement of tests and reagents ... SARS-CoV-2 diagnostic tests have become an important tool for pandemic control ... recognized the sequence TFPPTEPKKDKKK located at the C-terminus of the N protein of main SARS-CoV-2 variants ...

Abstract	SARS-CoV-2 diagnostic tests have become an important tool for pandemic control. Among the alternatives for COVID-19 diagnosis, antigen rapid diagnostic tests (Ag-RDT) are very convenient and widely used. However, as SARS-CoV-2 variants may continuously emerge, the replacement of tests and reagents may be required to maintain the sensitivity of Ag-RDTs. Here, we describe the development and validation of an Ag-RDT during an outbreak of the Omicron variant, including the characterization of a new monoclonal antibody (anti-DTC-N 1B3 mAb) that recognizes the Nucleocapsid protein (N). The anti-DTC-N 1B3 mAb recognized the sequence TFPPTEPKKDKKK located at the C-terminus of the N protein of main SARS-CoV-2 variants of concern. Accordingly, the Ag-RDT prototypes using the anti-DTC-N 1B3 mAB detected all the SARS-CoV-2 variants-Wuhan, Alpha, Gamma, Delta, P2 and Omicron. The performance of the best prototype (sensitivity of 95.2% for samples with Ct ≤ 25; specificity of 98.3% and overall accuracy of 85.0%) met the WHO recommendations. Moreover, results from a patients' follow-up study indicated that, if performed within the first three days after onset of symptoms, the Ag-RDT displayed 100% sensitivity. Thus, the new mAb and the Ag-RDT developed herein may constitute alternative tools for COVID-19 point-of-care diagnosis and epidemiological surveillance.
Language	English
Publishing date	2023-09-28
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2720891-6
ISSN	2076-2607
ISSN	2076-2607
DOI	10.3390/microorganisms11102422
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Article ; Online: Development of a novel medium throughput flow-cytometry based micro-neutralisation test for SARS-CoV-2 with applications in clinical vaccine trials and antibody screening.

O'Reilly, Sophie / Kenny, Grace / Alrawahneh, Tamara / Francois, Nathan / Gu, Lili / Angeliadis, Matthew / de Masson d'Autume, Valentin / Garcia Leon, Alejandro / Feeney, Eoin R / Yousif, Obada / Cotter, Aoife / de Barra, Eoghan / Horgan, Mary / Mallon, Patrick W G / Gautier, Virginie

PloS one

2023 Volume 18, Issue 11, Page(s) e0294262

Abstract: Quantifying neutralising capacity of circulating SARS-COV-2 antibodies is critical in evaluating ... responses against live SARS-CoV-2 Wild Type and Variants of Concern (VOC) in convalescent/vaccinated ... SARS-CoV-2 Nucleoprotein intracellular staining. Half-maximal Neutralisation Titres (NT50) were ...

Abstract	Quantifying neutralising capacity of circulating SARS-COV-2 antibodies is critical in evaluating protective humoral immune responses generated post-infection/post-vaccination. Here we describe a novel medium-throughput flow cytometry-based micro-neutralisation test to evaluate Neutralising Antibody (NAb) responses against live SARS-CoV-2 Wild Type and Variants of Concern (VOC) in convalescent/vaccinated populations. Flow Cytometry-Based Micro-Neutralisation Test (Micro-NT) was performed in 96-well plates using clinical isolates WT-B, WT-B.1.177.18 and/or VOCs Beta and Omicron. Plasma samples (All Ireland Infectious Diseases (AIID) Cohort) were serially diluted (8 points, half-log) from 1:20 and pre-incubated with SARS-CoV-2 (1h, 37°C). Virus-plasma mixture were added onto Vero E6 or Vero E6/TMPRSS2 cells for 18h. Percentage infected cells was analysed by automated flow cytometry following trypsinisation, fixation and SARS-CoV-2 Nucleoprotein intracellular staining. Half-maximal Neutralisation Titres (NT50) were determined using non-linear regression. Our assay was compared to Plaque Reduction Neutralisation Test (PRNT) and validated against the First WHO International Standard for anti-SARS-CoV-2 immunoglobulin. Both Micro-NT and PRNT achieved comparable NT50 values. Further validation showed adequate correlation with PRNT using a panel of secondary standards of clinical convalescent and vaccinated plasma samples. We found the assay to be reproducible through measuring both repeatability and intermediate precision. Screening 190 convalescent samples and 11 COVID-19 naive controls (AIID cohort) we demonstrated that Micro-NT has broad dynamic range differentiating NT50s <1/20 to >1/5000. We could also characterise immune-escape VOC Beta and Omicron BA.5, achieving fold-reductions in neutralising capacity similar to those published. Our flow cytometry-based Micro-NT is a robust and reliable assay to quantify NAb titres, and has been selected as an endpoint in clinical trials.
MeSH term(s)	Humans ; Flow Cytometry ; SARS-CoV-2 ; Neutralization Tests ; COVID-19 ; Vaccines ; Antibodies, Neutralizing ; Antibodies, Viral
Chemical Substances	Vaccines ; Antibodies, Neutralizing ; Antibodies, Viral
Language	English
Publishing date	2023-11-30
Publishing country	United States
Document type	Journal Article
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0294262
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Article ; Online: Monoclonal antibody pairs against SARS-CoV-2 for rapid antigen test development.

Salcedo, Nol / Reddy, Ankita / Gomez, Adam R / Bosch, Irene / Herrera, Bobby Brooke

PLoS neglected tropical diseases

2022 Volume 16, Issue 3, Page(s) e0010311

Abstract: ... SARS-CoV-2 N during the acute phase of COVID-19. The rapid antigen tests developed in this study are ... of commercially available antigen tests to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2 ... flow chromatography. A candidate rapid antigen test for SARS-CoV-2 N was validated using nasal swab ...

Abstract	Background: The focus on laboratory-based diagnosis of coronavirus disease 2019 (COVID-19) warrants alternative public health tools such as rapid antigen tests. While there are a number of commercially available antigen tests to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), all cross-react with the genetically similar SARS-CoV-1 or require an instrument for results interpretation. Methodology/principal findings: We developed and validated rapid antigen tests that use pairs of murine-derived monoclonal antibodies (mAbs), along with gold nanoparticles, to detect SARS-CoV-2 with or without cross-reaction to SARS-CoV-1 and other coronaviruses. In this development, we demonstrate a robust antibody screening methodology for the selection of mAb pairs that can recognize SARS-CoV-2 spike (S) and nucleocapsid (N) proteins. Linear epitope mapping of the mAbs helped elucidate SARS-CoV-2 S and N interactions in lateral flow chromatography. A candidate rapid antigen test for SARS-CoV-2 N was validated using nasal swab specimens that were confirmed positive or negative by quantitative reverse-transcription polymerase chain reaction (RT-PCR). Test results were image-captured using a mobile phone and normalized signal pixel intensities were calculated; signal intensities were inversely correlated to RT-PCR cycle threshold (Ct) value. Conclusion/significance: Overall, our results suggest that the rapid antigen test is optimized to detect SARS-CoV-2 N during the acute phase of COVID-19. The rapid antigen tests developed in this study are alternative tools for wide scale public health surveillance of COVID-19.
MeSH term(s)	Animals ; Antibodies, Monoclonal ; COVID-19/diagnosis ; Gold ; Metal Nanoparticles ; Mice ; SARS-CoV-2 ; Sensitivity and Specificity
Chemical Substances	Antibodies, Monoclonal ; Gold (7440-57-5)
Language	English
Publishing date	2022-03-31
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2429704-5
ISSN	1935-2735 ; 1935-2735
ISSN (online)	1935-2735
ISSN	1935-2735
DOI	10.1371/journal.pntd.0010311
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Article ; Online: Evaluation of the analytical performance of anti-SARS-CoV-2 antibody test kits distributed or developed in Japan.

Shibata, Hiroko / Nishimura, Kazuko / Maeda, Takuya / Honma, Masamitsu / Goda, Yukihiro / Ishii-Watabe, Akiko / Saito, Yoshiro

Bioanalysis

2022 Volume 14, Issue 6, Page(s) 325–340

Abstract: Background: ...

Abstract	Background:
MeSH term(s)	Antibodies, Viral/blood ; Automation, Laboratory ; COVID-19 Serological Testing/instrumentation ; COVID-19 Serological Testing/methods ; COVID-19 Serological Testing/standards ; Enzyme-Linked Immunosorbent Assay/instrumentation ; Enzyme-Linked Immunosorbent Assay/methods ; Humans ; Immunoglobulin G/blood ; Japan ; Reagent Kits, Diagnostic ; SARS-CoV-2/immunology
Chemical Substances	Antibodies, Viral ; Immunoglobulin G ; Reagent Kits, Diagnostic
Language	English
Publishing date	2022-03-02
Publishing country	England
Document type	Evaluation Study ; Journal Article
ISSN	1757-6199
ISSN (online)	1757-6199
DOI	10.4155/bio-2021-0254
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Article ; Online: Development of a rapid neutralizing antibody test for SARS-CoV-2 and its application for neutralizing antibody screening and vaccinated serum testing.

Li, Yuchang / Wang, Mingyue / Wu, Hongzhen / Zhao, Hui / Dong, Lei / Li, Yunfei / Li, Xiaofeng / Tang, Ying / Zhang, Sen / Li, Jing / Qin, Chengfeng / Jiang, Tao / Deng, Yongqiang / Kang, Xiaoping

Infectious medicine

2022 Volume 1, Issue 2, Page(s) 95–102

Abstract: ... mutation frequency of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent, have ... with the inactivated SARS-CoV-2 vaccine.: Results: The sensitivity and specificity of the ACE2-Block ELISA were 96.3 ... the ACE2-S-RBD interaction. A plaque assay confirmed that ch-2C5 neutralized SARS-CoV-2, with NT ...

Abstract	Background: Since the outbreak of coronavirus disease (COVID-19), the high infection rate and mutation frequency of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent, have contributed to the ongoing global pandemic. Vaccination has become the most effective means of controlling COVID-19. Traditional neutralizing tests of sera are complex and labor-intensive, therefore, a rapid test for detecting neutralizing antibodies and antibody status post-immunization is needed. Methods: Based on the fact that antibodies exhibit neutralizing activity by blocking the binding of the S protein receptor-binding domain (S-RBD) to ACE2, we developed a rapid neutralizing antibody test, ACE2-Block-ELISA. To evaluate the sensitivity and specificity, we used 54 positive and 84 negative serum samples. We also tested the neutralizing activities of monoclonal antibodies (mAbs) and 214 sera samples from healthy individuals immunized with the inactivated SARS-CoV-2 vaccine. Results: The sensitivity and specificity of the ACE2-Block ELISA were 96.3% and 100%, respectively. For neutralizing mAb screening, ch-2C5 was selected for its ability to block the ACE2-S-RBD interaction. A plaque assay confirmed that ch-2C5 neutralized SARS-CoV-2, with NT Conclusions: This study developed and validated an ACE2-Block-ELISA to test the neutralizing activities of antibodies. As a rapid, inexpensive and easy-to-perform method, this ACE2-Block-ELISA has potential applications in rapid neutralizing mAb screening and SARS-CoV-2 vaccine evaluation.
Language	English
Publishing date	2022-04-26
Publishing country	China
Document type	Journal Article
ISSN	2772-431X
ISSN (online)	2772-431X
DOI	10.1016/j.imj.2022.04.003
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Article ; Online: Peptide microarray-based analysis of antibody responses to SARS-CoV-2 identifies unique epitopes with potential for diagnostic test development.

Holenya, Pavlo / Lange, Paul Joris / Reimer, Ulf / Woltersdorf, Wolfram / Panterodt, Thomas / Glas, Michael / Wasner, Mark / Eckey, Maren / Drosch, Michael / Hollidt, Jörg-Michael / Naumann, Michael / Kern, Florian / Wenschuh, Holger / Lange, Robert / Schnatbaum, Karsten / Bier, Frank F

European journal of immunology

2021 Volume 51, Issue 7, Page(s) 1839–1849

Abstract: Humoral immunity to the Severe Adult Respiratory Syndrome (SARS) Coronavirus (CoV)-2 is not fully ... between SARS-CoV-2 and other human coronaviruses (HCoVs) would have important implications for immune ... patient samples and n = 12 control samples were screened for antibodies against the entire SARS-CoV-2 ...

Abstract	Humoral immunity to the Severe Adult Respiratory Syndrome (SARS) Coronavirus (CoV)-2 is not fully understood yet but is a crucial factor of immune protection. The possibility of antibody cross-reactivity between SARS-CoV-2 and other human coronaviruses (HCoVs) would have important implications for immune protection but also for the development of specific diagnostic ELISA tests. Using peptide microarrays, n = 24 patient samples and n = 12 control samples were screened for antibodies against the entire SARS-CoV-2 proteome as well as the Spike (S), Nucleocapsid (N), VME1 (V), R1ab, and Protein 3a (AP3A) of the HCoV strains SARS, MERS, OC43, and 229E. While widespread cross-reactivity was revealed across several immunodominant regions of S and N, IgG binding to several SARS-CoV-2-derived peptides provided statistically significant discrimination between COVID-19 patients and controls. Selected target peptides may serve as capture antigens for future, highly COVID-19-specific diagnostic antibody tests.
MeSH term(s)	Adult ; Aged ; Amino Acid Sequence/genetics ; Antibodies, Viral/blood ; Antibodies, Viral/immunology ; COVID-19/diagnosis ; Coronavirus 229E, Human/immunology ; Coronavirus Nucleocapsid Proteins/immunology ; Coronavirus OC43, Human/immunology ; Cross Reactions/immunology ; Diagnostic Tests, Routine ; Female ; Humans ; Immunoglobulin G/blood ; Immunoglobulin G/immunology ; Male ; Middle Aged ; Middle East Respiratory Syndrome Coronavirus/immunology ; Phosphoproteins/immunology ; Protein Array Analysis/methods ; Proteome/immunology ; SARS Virus/immunology ; SARS-CoV-2/immunology ; Spike Glycoprotein, Coronavirus/immunology ; Viral Proteins/immunology ; Young Adult
Chemical Substances	Antibodies, Viral ; Coronavirus Nucleocapsid Proteins ; Immunoglobulin G ; Phosphoproteins ; Proteome ; Spike Glycoprotein, Coronavirus ; Viral Proteins ; nucleocapsid phosphoprotein, SARS-CoV-2 ; spike protein, SARS-CoV-2
Language	English
Publishing date	2021-05-07
Publishing country	Germany
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	120108-6
ISSN	1521-4141 ; 0014-2980
ISSN (online)	1521-4141
ISSN	0014-2980
DOI	10.1002/eji.202049101
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Article: Evaluation of diagnostic accuracy of developed rapid SARS-COV-2 IgG antibody test kit using novel diluent system.

Maya, Vani / Kaviyil, Jyothi Embekkat / Ponnambath, Dinoop Koral / Nair, Renjith P / Bhatt, Anugya / Pilankatta, Rajendra / Valliyot, Lathika / Dungdung, Ranjeet / Veettil, Ramdas Athikkal / Gopi, Manoj

Virusdisease

2021 Volume 32, Issue 1, Page(s) 78–84

Abstract: ... was to develop a rapid test kit for SARS-COV-2 with a novel diluent system to enhance the efficacy ... against SARS-COV-2 virus peptides were analyzed using 25 positive and 25 negative confirmed clinical ... of SARS-COV-2 infection and additionally, for effective monitoring of convalescent sera therapy. ...

Abstract	Immunochromatographic assay kits are used in primary diagnostics which is based on the principle of antigen and antibody interaction. These kits play pivotal role in rapid surveillance of infectious diseases at early stages as well as for the surveillance of the contagious diseases. The immunochromatographic test kits lacks sensitivity and specificity with certain diseases. In this study, our intention was to develop a rapid test kit for SARS-COV-2 with a novel diluent system to enhance the efficacy of antigen-antibody binding and thereby the improvement in the sensitivity outlined. Finally, IgG antibodies against SARS-COV-2 virus peptides were analyzed using 25 positive and 25 negative confirmed clinical samples. The sensitivity of the clinical studies showed 91% sensitivity and 100% specificity. Therefore, the authors propose that this assay will be a potential tool for efficient community or sentinel surveillance of SARS-COV-2 infection and additionally, for effective monitoring of convalescent sera therapy.
Language	English
Publishing date	2021-03-04
Publishing country	India
Document type	Journal Article
ZDB-ID	2846993-8
ISSN	2347-3517 ; 2347-3584
ISSN (online)	2347-3517
ISSN	2347-3584
DOI	10.1007/s13337-021-00669-4
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