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Article ; Online: A magnetic bead immunoassay to detect high affinity human IgG reactive to SARS-CoV-2 Spike S1 RBD produced in Escherichia coli.

Conzentino, Marcelo S / Gonçalves, Ana C A / Paula, Nigella M / Rego, Fabiane G M / Zanette, Dalila L / Aoki, Mateus N / Nardin, Jeanine M / Huergo, Luciano Fernandes

Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology

2022 Volume 53, Issue 3, Page(s) 1263–1269

Abstract: Immunological assays to detect SARS-CoV-2 Spike Receptor Binding Domain (RBD) antigen seroconversion in humans are important tools to monitor the levels of protecting antibodies in the population in response to infection and/or immunization. Here we ... ...

Abstract	Immunological assays to detect SARS-CoV-2 Spike Receptor Binding Domain (RBD) antigen seroconversion in humans are important tools to monitor the levels of protecting antibodies in the population in response to infection and/or immunization. Here we describe a simple, low cost, and high throughput Ni
MeSH term(s)	Antibodies, Viral ; COVID-19 ; Escherichia coli/genetics ; Humans ; Immunoassay/methods ; Immunoglobulin G ; Magnetic Phenomena ; SARS-CoV-2/genetics ; Spike Glycoprotein, Coronavirus/genetics
Chemical Substances	Antibodies, Viral ; Immunoglobulin G ; Spike Glycoprotein, Coronavirus ; spike protein, SARS-CoV-2
Language	English
Publishing date	2022-04-14
Publishing country	Brazil
Document type	Journal Article
ZDB-ID	2017175-4
ISSN	1678-4405 ; 1517-8382
ISSN (online)	1678-4405
ISSN	1517-8382
DOI	10.1007/s42770-022-00753-x
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Article: Population-Based Analysis of the Immunoglobulin G Response to Different COVID-19 Vaccines in Brazil.

Paula, Nigella M / Conzentino, Marcelo S / Gonçalves, Ana C A / da Silva, Renata / Weissheimer, Karin V / Kluge, Carlos H S / Marins, Paulo H S A / Camargo, Haxley S C / Farias, Lucas R P / Sant'Ana, Thamyres P / Vargas, Letícia R / Aldrighi, Juliane D / Lima, Ênio S / Jacotenski, Guiomar T / Pedrosa, Fabio O / Gonçalves, Alan G / Joucoski, Emerson / Huergo, Luciano F

Vaccines

2022 Volume 11, Issue 1

Abstract: ... ChAdOx1-S (AstraZeneca-University of Oxford) and BNT162b2 (Pfizer-BioNTech) vaccines. The titers of IgG ... in individuals vaccinated with BNT162b2, followed by ChAdOx1-S and CoronaVac. The anti-spike IgG response was ... infections boosted anti-spike IgG in those individuals who completed primary vaccination with ChAdOx1-S and ...

Abstract	(1) Background: COVID-19 vaccination in Brazil has been performed mostly with CoronaVac (Sinovac), ChAdOx1-S (AstraZeneca-University of Oxford) and BNT162b2 (Pfizer-BioNTech) vaccines. The titers of IgG antibodies reactive to the SARS-CoV-2 spike protein correlate with vaccine efficacy. Studies comparing vaccine immunogenicity in a real-world scenario are lacking. (2) Methods: We performed a population-based study to analyze the immunoglobulin G response to different COVID-19 vaccines. Citizens older than 18 years (n = 2376) provided personal data, a self-declaration of any previous COVID-19 positive tests and information regarding COVID-19 vaccination: the vaccine popular name and the date of each dose. Blood samples were collected and the levels of IgG reactive to SARS-CoV-2 antigens were determined and compared between different vaccine groups. (3) Results: The seroconversion for anti-spike IgG achieved > 95% by February 2022 and maintained stable until June 2022. Higher anti-spike IgG titers were detected in individuals vaccinated with BNT162b2, followed by ChAdOx1-S and CoronaVac. The anti-spike IgG response was negatively correlated with age and interval after the second dose for the BNT162b2 vaccine. Natural infections boosted anti-spike IgG in those individuals who completed primary vaccination with ChAdOx1-S and CoronaVac, but not with BNT162b2. The levels of anti-spike IgG increased with the number of vaccine doses administered. The application of BNT162b2 as a 3rd booster dose resulted in high anti-spike IgG antibody titers, despite the type of vaccine used during primary vaccination. (4) Conclusions: Our data confirmed the effectiveness of the Brazilian vaccination program. Of the vaccines used in Brazil, BNT162b2 performed better to elicit anti-spike protein IgG after primary vaccination and as a booster dose and thus should be recommended as a booster whenever available. A continuous COVID-19 vaccination program will be required to sustain anti-spike IgG antibodies in the population.
Language	English
Publishing date	2022-12-22
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2703319-3
ISSN	2076-393X
ISSN	2076-393X
DOI	10.3390/vaccines11010021
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Article ; Online: SARS-CoV-2 Seroconversion in Response to Infection and Vaccination: a Time Series Local Study in Brazil.

Huergo, Luciano F / Paula, Nigella M / Gonçalves, Ana C A / Kluge, Carlos H S / Marins, Paulo H S A / Camargo, Haxley S C / Sant'Ana, Thamyres P / Farias, Lucas R P / Aldrighi, Juliane D / Lima, Ênio S / Jacotenski, Guiomar T / Vargas, Letícia R / Costa, Gisele / Weissheimer, Karin V / Nazário, Maria G / Teixeira, Kádima N / Conzentino, Marcelo S

Microbiology spectrum

2022 Volume 10, Issue 4, Page(s) e0102622

Abstract: The investigation of antibodies raised against different severe acute respiratory syndrome coronavirus (SARS-CoV-2) antigens can help to determine the extent of previous SARS-CoV-2 infections in the population and track the humoral response to ... ...

Abstract	The investigation of antibodies raised against different severe acute respiratory syndrome coronavirus (SARS-CoV-2) antigens can help to determine the extent of previous SARS-CoV-2 infections in the population and track the humoral response to vaccination. Therefore, serological surveys can provide key information to better manage the pandemic and/or to implement the most effective vaccination program. Here we describe a time series anti-nucleocapsid, anti-spike IgG serological survey analysis in the city of Matinhos, PR, Brazil during the year of 2021. Seroconversion rates to the nucleocapsid antigen were not influenced by gender or age. The serological data support that the coronavirus disease 2019 (COVID-19) infection rate is ~50% higher than official numbers. Furthermore, by applying serological data, the corrected infection fatality rate was estimated to be lower than 2.4% in contrast with the official estimative of 3.6%. The rates of IgG reactive to spike antigen resembled the curve of the fraction the population that had taken the second vaccine dose. Up to 82% of spike seroconversion was detected in the end of 2021, confirming the effectiveness of the COVID-19 vaccination program in the city. This SARS-CoV-2 serological study unraveled the SARS-CoV-2 infection rates and the response to vaccination in the city of Matinhos.
MeSH term(s)	Antibodies, Viral ; Brazil/epidemiology ; COVID-19/epidemiology ; COVID-19/prevention & control ; COVID-19 Vaccines ; Humans ; Immunoglobulin G ; SARS-CoV-2 ; Seroconversion ; Spike Glycoprotein, Coronavirus ; Time Factors ; Vaccination
Chemical Substances	Antibodies, Viral ; COVID-19 Vaccines ; Immunoglobulin G ; Spike Glycoprotein, Coronavirus ; spike protein, SARS-CoV-2
Language	English
Publishing date	2022-06-30
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2807133-5
ISSN	2165-0497 ; 2165-0497
ISSN (online)	2165-0497
ISSN	2165-0497
DOI	10.1128/spectrum.01026-22
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Article ; Online: Ultra-fast, high throughput and inexpensive detection of SARS-CoV-2 seroconversion using Ni

Conzentino, Marcelo S / Santos, Tatielle P C / Selim, Khaled A / Wagner, Berenike / Alford, Janette T / Deobald, Nelli / Paula, Nigela M / Rego, Fabiane G M / Zanette, Dalila L / Aoki, Mateus N / Nardin, Jeanine M / Huergo, Maria C C / Reis, Rodrigo A / Huergo, Luciano F

Analytical biochemistry

2021 Volume 631, Page(s) 114360

Abstract: To monitor the levels of protecting antibodies raised in the population in response to infection and/or to immunization with SARS-CoV-2, we need a technique that allows high throughput and low-cost quantitative analysis of human IgG antibodies reactive ... ...

Abstract	To monitor the levels of protecting antibodies raised in the population in response to infection and/or to immunization with SARS-CoV-2, we need a technique that allows high throughput and low-cost quantitative analysis of human IgG antibodies reactive against viral antigens. Here we describe an ultra-fast, high throughput and inexpensive assay to detect SARS-CoV-2 seroconversion in humans. The assay is based on Ni
MeSH term(s)	Antibodies, Viral/blood ; Antibodies, Viral/immunology ; Antigens, Viral/blood ; Antigens, Viral/immunology ; COVID-19/blood ; COVID-19/diagnosis ; COVID-19/immunology ; COVID-19 Serological Testing/economics ; COVID-19 Serological Testing/methods ; Enzyme-Linked Immunosorbent Assay/economics ; Enzyme-Linked Immunosorbent Assay/methods ; Humans ; Immunoglobulin G/blood ; Immunoglobulin G/immunology ; Magnets/chemistry ; Nickel/chemistry ; SARS-CoV-2/immunology ; SARS-CoV-2/isolation & purification ; Sensitivity and Specificity ; Seroconversion ; Time Factors
Chemical Substances	Antibodies, Viral ; Antigens, Viral ; Immunoglobulin G ; Nickel (7OV03QG267)
Language	English
Publishing date	2021-09-03
Publishing country	United States
Document type	Evaluation Study ; Journal Article
ZDB-ID	1110-1
ISSN	1096-0309 ; 0003-2697
ISSN (online)	1096-0309
ISSN	0003-2697
DOI	10.1016/j.ab.2021.114360
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Article ; Online: Populational analysis of the immunoglobulin G response to different COVID-19 vaccines in Brazil

Paula, Nigella M / Conzentino, Marcelo dos Santos / Goncalves, Ana C / Silva, Renata / Weissheimer, Karin V / Kluge, Carlos H / Marins, Paulo H / Camargo, Haxely S / Farias, Lucas R / Santana, Thamyres P / Vargas, Leticia R / Aldrighi, Juliane D / Lima, Enio S / Jacotenski, Guiomar T / Goncalves, Alan G / Joucoski, Emerson / Huergo, Luciano

medRxiv

Abstract: Vaccination is a strategy that confers protection against symptomatic infections and/or development of severe COVID-19. In Brazil, COVID-19 vaccination began in January 2021 and has been performed using vaccines from different manufactures including ... ...

Abstract	Vaccination is a strategy that confers protection against symptomatic infections and/or development of severe COVID-19. In Brazil, COVID-19 vaccination began in January 2021 and has been performed using vaccines from different manufactures including CoronaVac (Sinovac), ChAdOx1 (Oxford/AstraZeneca) and BNT162b2 (Pfizer/BioNTech). One of the main protective mechanisms triggered by vaccination involves the production of IgG antibodies reactive to the Spike antigen of SARS-CoV-2, the levels of which correlates with vaccine efficacy. Although phase III clinical studies confirmed the efficacy of the vaccines used in Brazil, there are just few studies comparing vaccine immunogenicity in a real-world scenario. This study aimed to depict the IgG response to natural infections and to vaccination using different types of vaccines at population scale in Matinhos, a city located in south of Brazil. Nucleocapsid seroconversion rates indicated that more than a quarter of the cohort has been subjected to natural infections by SARS-CoV-2 by the first trimester of 2022. Spike seroconversion rates achieved >95% by February 2022 and maintained stable as far as June 2022 confirming the effectiveness of the vaccination program. Immunogenicity concerning IgG reactive to Spike was higher using the BNT162b2 vaccine, followed by ChAdOx1 and CoronaVac. Natural infections boosted IgG levels reactive to Spike in those individuals that completed primary vaccination with ChAdOx1 and CoronaVac but not with BNT162b2. The levels of IgG reactive to Spike increased with the number of vaccine doses administered. The application of BNT162b2 as booster dose resulted in high levels of IgG reactive to Spike which were similar despite the type of the vaccine used during primary vaccination.
Keywords	covid19
Language	English
Publishing date	2022-09-29
Publisher	Cold Spring Harbor Laboratory Press
Document type	Article ; Online
DOI	10.1101/2022.09.28.22280449
Database	COVID19

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Article: Ultra-fast, high throughput and inexpensive detection of SARS-CoV-2 seroconversion using Ni2+ magnetic beads

Conzentino, Marcelo S. / Santos, Tatielle P.C. / Selim, Khaled A. / Wagner, Berenike / Alford, Janette T. / Deobald, Nelli / Paula, Nigela M. / Rego, Fabiane G.M. / Zanette, Dalila L. / Aoki, Mateus N. / Nardin, Jeanine M. / Huergo, Maria C.C. / Reis, Rodrigo A. / Huergo, Luciano F.

Analytical biochemistry. 2021 Oct. 15, v. 631

2021

Abstract	To monitor the levels of protecting antibodies raised in the population in response to infection and/or to immunization with SARS-CoV-2, we need a technique that allows high throughput and low-cost quantitative analysis of human IgG antibodies reactive against viral antigens. Here we describe an ultra-fast, high throughput and inexpensive assay to detect SARS-CoV-2 seroconversion in humans. The assay is based on Ni²⁺ magnetic particles coated with His tagged SARS-CoV-2 antigens. A simple and inexpensive 96 well plate magnetic extraction/homogenization process is described which allows the simultaneous analysis of 96 samples and delivers results in 7 min with high accuracy.
Keywords	Severe acute respiratory syndrome coronavirus 2 ; homogenization ; humans ; immunization ; magnetism ; quantitative analysis ; seroconversion
Language	English
Dates of publication	2021-1015
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	1110-1
ISSN	1096-0309 ; 0003-2697
ISSN (online)	1096-0309
ISSN	0003-2697
DOI	10.1016/j.ab.2021.114360
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Antigen production and development of an indirect ELISA based on the nucleocapsid protein to detect human SARS-CoV-2 seroconversion.

Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology

2021 Volume 52, Issue 4, Page(s) 2069–2073

Abstract: Serological assays are important tools to identify previous exposure to SARS-CoV-2, helping to track COVID-19 cases and determine the level of humoral response to SARS-CoV-2 infections and/or immunization to future vaccines. Here, the SARS-CoV-2 ... ...

Abstract	Serological assays are important tools to identify previous exposure to SARS-CoV-2, helping to track COVID-19 cases and determine the level of humoral response to SARS-CoV-2 infections and/or immunization to future vaccines. Here, the SARS-CoV-2 nucleocapsid protein was expressed in Escherichia coli and purified to homogeneity and high yield using a single chromatography step. The purified SARS-CoV-2 nucleocapsid protein was used to develop an indirect enzyme-linked immunosorbent assay for the identification of human SARS-CoV-2 seroconverts. The assay sensitivity and specificity were determined analyzing sera from 140 RT-qPCR-confirmed COVID-19 cases and 210 pre-pandemic controls. The assay operated with 90% sensitivity and 98% specificity; identical accuracies were obtained in head-to-head comparison with a commercial ELISA kit. Antigen-coated plates were stable for up to 3 months at 4 °C. The ELISA method described is ready for mass production and will be an additional tool to track COVID-19 cases.
MeSH term(s)	Antibodies, Viral/blood ; COVID-19/diagnosis ; COVID-19/immunology ; Coronavirus Nucleocapsid Proteins/immunology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunity, Humoral ; Nucleocapsid Proteins/genetics ; Phosphoproteins/immunology ; Sensitivity and Specificity ; Seroconversion
Chemical Substances	Antibodies, Viral ; Coronavirus Nucleocapsid Proteins ; Nucleocapsid Proteins ; Phosphoproteins ; nucleocapsid phosphoprotein, SARS-CoV-2
Language	English
Publishing date	2021-08-03
Publishing country	Brazil
Document type	Journal Article
ZDB-ID	2017175-4
ISSN	1678-4405 ; 1517-8382
ISSN (online)	1678-4405
ISSN	1517-8382
DOI	10.1007/s42770-021-00556-6
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Article ; Online: SARS-CoV-2 seroconversion in response to infection and vaccination: A time series local study in Brazil

Huergo, Luciano / Paula, Nigella M / Goncalves, Ana C.A. / Kluge, Carlos H.S / Martins, Paulo H.S.A / Camargo, Haxley S.C / Santana, Thamyres P / Farias, Lucas R.P. / Lima, Enio S / Aldrighi, Juliane D. / Jacotenski, Guiomar T / Vargas, Leticia R / Costa, Gisele / Weissheimer, Karin V / Nazario, Mariana G / Teixeira, Kadima N / Conzentino, Marcelo dos Santos

medRxiv

Abstract: The investigation of antibodies raised against different SARS-CoV-2 antigens can help to determine the extent of previous SARS-CoV-2 infections in the population and track the humoral response to vaccination. Therefore, serological surveys can provide ... ...

Abstract	The investigation of antibodies raised against different SARS-CoV-2 antigens can help to determine the extent of previous SARS-CoV-2 infections in the population and track the humoral response to vaccination. Therefore, serological surveys can provide key information to better manage the pandemic and/or to implement the most effective vaccination program. Here we describe a time series anti-Nucleocapsid, anti-Spike IgG serological survey analysis in the city of Matinhos, PR, Brazil during the year of 2021. Seroconversion rates to the Nucleocapsid antigen was not influenced by gender or age. Comparison of the serological data with official COVID-19 cases in the city suggest that case sub notification is higher than 47%. Furthermore, by applying serological data, the corrected infection fatality rate was estimated to be lower than 2.4 % in contrast with the official estimative of 3.6 %. The rates of IgG reactive to Spike antigen resembled the curve of the fraction the population that had taken the second vaccine dose. Up to 82% of Spike seroconversion was detected in the end of 2021 confirming the effective of the COVID-19 vaccination program in the city. This SARS-CoV-2 serological study unraveled the SARS-CoV-2 infection rates and the response to vaccination in the city of Matinhos, it is likely that the numbers reported here may be similar in other cities in Brazil.
Keywords	covid19
Language	English
Publishing date	2022-03-12
Publisher	Cold Spring Harbor Laboratory Press
Document type	Article ; Online
DOI	10.1101/2022.03.10.22271805
Database	COVID19

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Article ; Online: Magnetic bead-based ELISA allow inexpensive, rapid and quantitative detection of human antibodies against SARS-CoV-2

Huergo, Luciano F / Conzentino, Marcelo S / Gerhardt, Edileusa C M / Santos, Adrian R.S. / Pedrosa, Fabio de Oliveira / Souza, Emanuel M / Nogueira, Meri B / Forchhammer, Karl / Rego, Fabiane G.M / Raboni, Sonia M / Reis, Rodrigo A

medRxiv

Abstract: Here we describe a novel immunogenic method to detect COVID-19. The method is a chromogenic magnetic bead-based ELISA which allows inexpensive and quantitative detection of human IgG or IgM antibodies against SARS-CoV-2 in serum or whole blood samples in ...

Abstract	Here we describe a novel immunogenic method to detect COVID-19. The method is a chromogenic magnetic bead-based ELISA which allows inexpensive and quantitative detection of human IgG or IgM antibodies against SARS-CoV-2 in serum or whole blood samples in just 12 minutes. As a proof of concept, we compared the performance of our new method to classical ELISA. Person correlation between optical densities obtained using the two methods was 0.98. The novel magnetic bead-based ELISA performed better than classic ELISA to discriminate one COVID-19 case carrying low antibody titer. The chromogenic magnetic bead-based ELISA method described here can be applied in formats for both point of care and high throughput analysis. The method is readily adaptable to use other protein-based antigens and is readily adaptable to investigate other diseases and other applications.
Keywords	covid19
Language	English
Publishing date	2020-07-29
Publisher	Cold Spring Harbor Laboratory Press
Document type	Article ; Online
DOI	10.1101/2020.07.26.20162255
Database	COVID19

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Article ; Online: Ultra-fast, high throughput and inexpensive detection of SARS-CoV-2 seroconversion

Conzentino, Marcelo S / Santos, Tatiele P.C. / Selim, Khaled / Wagner, Berenike / Alford, Janette T / Deobald, Nelli / Paula, Nigella M / Rego, Fabiane G.M / Zanette, Dalila L / Aoki, Mateus N / Nardin, Jeanine M / Huergo, Maria C.C / Reis, Rodrigo A / Huergo, Luciano

medRxiv

Abstract: A technique allowing high throughput, fast and low-cost quantitative analysis of human IgG antibodies reacting to SARS-CoV-2 antigens will be required to understand the levels of protecting antibodies in the population raised in response to infections ... ...

Abstract	A technique allowing high throughput, fast and low-cost quantitative analysis of human IgG antibodies reacting to SARS-CoV-2 antigens will be required to understand the levels of protecting antibodies in the population raised in response to infections and/or to immunization. We described previously a fast, simple, and inexpensive Ni2+ magnetic bead immunoassay which allowed detection of human antibodies reacting against the SARS-CoV-2 nucleocapsid protein using a minimal amount of serum or blood. A major draw-back of the previously described system was that it only processed 12 samples simultaneously. Here we describe a manually operating inexpensive 96 well plate magnetic extraction / homogenization process which allows high throughput analysis delivering results of 96 samples in chromogenic format in 12 minutes or in fluorescent ultrafast format which takes only 7 minutes. We also show that His tag antigen purification can be performed on the fly while loading antigens to the Ni2+ magnetic beads in a process which takes only 12 min reducing the pre analytical time and cost. Finally, we show that the magnetic bead immunoassay is antigen flexible and can be performed using either Nucleocapsid, Spike or Spike RBD. The method performed with low inter and intra assay variability using different antigens and detection modes and was able to deliver >99.5% specificity and >95% sensitivity for a cohort of 203 pre pandemic and 63 COVID-19 positive samples.
Keywords	covid19
Language	English
Publishing date	2021-04-19
Publisher	Cold Spring Harbor Laboratory Press
Document type	Article ; Online
DOI	10.1101/2021.04.13.21255396
Database	COVID19

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