A magnetic bead immunoassay to detect high affinity human IgG reactive to SARS-CoV-2 Spike S1 RBD produced in Escherichia coli
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 N...
Gespeichert in:
| Veröffentlicht in: | Brazilian journal of microbiology 2022-09, Vol.53 (3), p.1263-1269 |
|---|---|
| Hauptverfasser: | , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 1269 |
|---|---|
| container_issue | 3 |
| container_start_page | 1263 |
| container_title | Brazilian journal of microbiology |
| container_volume | 53 |
| creator | 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 |
| description | 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
2+
magnetic bead immunoassay to detect human IgG reactive to Spike S1 RBD Receptor Binding Domain produced in
Escherichia coli.
A 6xHis-tagged Spike S1 RBD was expressed in
E. coli
and purified by affinity chromatography. The protein was mobilized on the surface of Ni
2+
magnetic beads and used to investigate the presence of reactive IgG in the serum obtained from pre-pandemic and COVID-19 confirmed cases. The method was validated with a cohort of 290 samples and an area under the receiver operating characteristic curve of 0.94 was obtained. The method was operated with > 82% sensitivity at 98% specificity and was also able to track human IgG raised in response to vaccination with Comirnaty at > 85% sensitivity. The IgG signal obtained with the described method was well-correlated with the signal obtained when pre fusion Spike produced in HEK cell lines was used as antigen. This novel low-cost and high throughput immunoassay may act as an important tool to investigate protecting IgG antibodies against SARS-CoV-2 in the human population. |
| doi_str_mv | 10.1007/s42770-022-00753-x |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_9009495</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2651693605</sourcerecordid><originalsourceid>FETCH-LOGICAL-c474t-e428d5a0c21484e24e5238f8f4484bd0ff89b424c5259c93bd2540f7c40106063</originalsourceid><addsrcrecordid>eNp9kU1v1DAQhiMEoqXwBzggS1y4GCaOnTgXpGUppVIlpC5wtRxnkrgk9tZOqu6F346XLeXjwMkezTOvPXqy7HkOr3OA6k3krKqAAmM0laKgtw-y47ysJOUcxMN0F3lFZSHZUfYkxisAJoCzx9lRITgroZTH2fcVmXTvcLaGNKhbYqdpcV7HqHdk9qTFGc1MBtsPRHeddXbekWGZtCPn_RkJqM1sb3CPblaXG7r2Xykjm639hmSTk8t378k2-HYxmKIdOY1mwGDNYDUxfrRPs0edHiM-uztPsi8fTj-vP9KLT2fn69UFNbziM0XOZCs0GJZzyZFxFKyQnex4KpsWuk7WDWfcCCZqUxdNywSHrjIcckiLFifZ20PudmkmbA26OehRbYOddNgpr636u-PsoHp_o2qAmtciBby6Cwj-esE4q8lGg-OoHfolKlaKvKyLEvboy3_QK78El9ZTrAJZlUlOkSh2oEzwMQbs7j-Tg9rrVQe9KulVP_Wq2zT04s817kd--UxAcQBiarkew--3_xP7A50ur6U</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2708764053</pqid></control><display><type>article</type><title>A magnetic bead immunoassay to detect high affinity human IgG reactive to SARS-CoV-2 Spike S1 RBD produced in Escherichia coli</title><source>SpringerLink (Online service)</source><source>PubMed Central</source><source>Alma/SFX Local Collection</source><creator>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</creator><creatorcontrib>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</creatorcontrib><description>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
2+
magnetic bead immunoassay to detect human IgG reactive to Spike S1 RBD Receptor Binding Domain produced in
Escherichia coli.
A 6xHis-tagged Spike S1 RBD was expressed in
E. coli
and purified by affinity chromatography. The protein was mobilized on the surface of Ni
2+
magnetic beads and used to investigate the presence of reactive IgG in the serum obtained from pre-pandemic and COVID-19 confirmed cases. The method was validated with a cohort of 290 samples and an area under the receiver operating characteristic curve of 0.94 was obtained. The method was operated with > 82% sensitivity at 98% specificity and was also able to track human IgG raised in response to vaccination with Comirnaty at > 85% sensitivity. The IgG signal obtained with the described method was well-correlated with the signal obtained when pre fusion Spike produced in HEK cell lines was used as antigen. This novel low-cost and high throughput immunoassay may act as an important tool to investigate protecting IgG antibodies against SARS-CoV-2 in the human population.</description><identifier>ISSN: 1517-8382</identifier><identifier>EISSN: 1678-4405</identifier><identifier>DOI: 10.1007/s42770-022-00753-x</identifier><identifier>PMID: 35426068</identifier><language>eng</language><publisher>Cham: Springer International Publishing</publisher><subject>Affinity ; Affinity chromatography ; Antibodies ; Antigens ; Beads ; Binding ; Biomedical and Life Sciences ; Cell lines ; Clinical Microbiology - Research Paper ; COVID-19 ; Domains ; E coli ; Escherichia coli ; Food Microbiology ; Human populations ; Immunization ; Immunoassay ; Immunoglobulin G ; Immunology ; Life Sciences ; Low cost ; Medical Microbiology ; Microbial Ecology ; Microbial Genetics and Genomics ; Microbiology ; Mycology ; Pandemics ; Public health ; Receptors ; Sensitivity ; Seroconversion ; Severe acute respiratory syndrome ; Severe acute respiratory syndrome coronavirus 2 ; Vaccination ; Vaccines ; Viral diseases</subject><ispartof>Brazilian journal of microbiology, 2022-09, Vol.53 (3), p.1263-1269</ispartof><rights>The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia 2022</rights><rights>2022. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.</rights><rights>The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia 2022.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c474t-e428d5a0c21484e24e5238f8f4484bd0ff89b424c5259c93bd2540f7c40106063</citedby><cites>FETCH-LOGICAL-c474t-e428d5a0c21484e24e5238f8f4484bd0ff89b424c5259c93bd2540f7c40106063</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9009495/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC9009495/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,881,27901,27902,41464,42533,51294,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/35426068$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Conzentino, Marcelo S.</creatorcontrib><creatorcontrib>Gonçalves, Ana C. A.</creatorcontrib><creatorcontrib>Paula, Nigella M.</creatorcontrib><creatorcontrib>Rego, Fabiane G. M.</creatorcontrib><creatorcontrib>Zanette, Dalila L.</creatorcontrib><creatorcontrib>Aoki, Mateus N.</creatorcontrib><creatorcontrib>Nardin, Jeanine M.</creatorcontrib><creatorcontrib>Huergo, Luciano Fernandes</creatorcontrib><title>A magnetic bead immunoassay to detect high affinity human IgG reactive to SARS-CoV-2 Spike S1 RBD produced in Escherichia coli</title><title>Brazilian journal of microbiology</title><addtitle>Braz J Microbiol</addtitle><addtitle>Braz J Microbiol</addtitle><description>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
2+
magnetic bead immunoassay to detect human IgG reactive to Spike S1 RBD Receptor Binding Domain produced in
Escherichia coli.
A 6xHis-tagged Spike S1 RBD was expressed in
E. coli
and purified by affinity chromatography. The protein was mobilized on the surface of Ni
2+
magnetic beads and used to investigate the presence of reactive IgG in the serum obtained from pre-pandemic and COVID-19 confirmed cases. The method was validated with a cohort of 290 samples and an area under the receiver operating characteristic curve of 0.94 was obtained. The method was operated with > 82% sensitivity at 98% specificity and was also able to track human IgG raised in response to vaccination with Comirnaty at > 85% sensitivity. The IgG signal obtained with the described method was well-correlated with the signal obtained when pre fusion Spike produced in HEK cell lines was used as antigen. This novel low-cost and high throughput immunoassay may act as an important tool to investigate protecting IgG antibodies against SARS-CoV-2 in the human population.</description><subject>Affinity</subject><subject>Affinity chromatography</subject><subject>Antibodies</subject><subject>Antigens</subject><subject>Beads</subject><subject>Binding</subject><subject>Biomedical and Life Sciences</subject><subject>Cell lines</subject><subject>Clinical Microbiology - Research Paper</subject><subject>COVID-19</subject><subject>Domains</subject><subject>E coli</subject><subject>Escherichia coli</subject><subject>Food Microbiology</subject><subject>Human populations</subject><subject>Immunization</subject><subject>Immunoassay</subject><subject>Immunoglobulin G</subject><subject>Immunology</subject><subject>Life Sciences</subject><subject>Low cost</subject><subject>Medical Microbiology</subject><subject>Microbial Ecology</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbiology</subject><subject>Mycology</subject><subject>Pandemics</subject><subject>Public health</subject><subject>Receptors</subject><subject>Sensitivity</subject><subject>Seroconversion</subject><subject>Severe acute respiratory syndrome</subject><subject>Severe acute respiratory syndrome coronavirus 2</subject><subject>Vaccination</subject><subject>Vaccines</subject><subject>Viral diseases</subject><issn>1517-8382</issn><issn>1678-4405</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><recordid>eNp9kU1v1DAQhiMEoqXwBzggS1y4GCaOnTgXpGUppVIlpC5wtRxnkrgk9tZOqu6F346XLeXjwMkezTOvPXqy7HkOr3OA6k3krKqAAmM0laKgtw-y47ysJOUcxMN0F3lFZSHZUfYkxisAJoCzx9lRITgroZTH2fcVmXTvcLaGNKhbYqdpcV7HqHdk9qTFGc1MBtsPRHeddXbekWGZtCPn_RkJqM1sb3CPblaXG7r2Xykjm639hmSTk8t378k2-HYxmKIdOY1mwGDNYDUxfrRPs0edHiM-uztPsi8fTj-vP9KLT2fn69UFNbziM0XOZCs0GJZzyZFxFKyQnex4KpsWuk7WDWfcCCZqUxdNywSHrjIcckiLFifZ20PudmkmbA26OehRbYOddNgpr636u-PsoHp_o2qAmtciBby6Cwj-esE4q8lGg-OoHfolKlaKvKyLEvboy3_QK78El9ZTrAJZlUlOkSh2oEzwMQbs7j-Tg9rrVQe9KulVP_Wq2zT04s817kd--UxAcQBiarkew--3_xP7A50ur6U</recordid><startdate>20220901</startdate><enddate>20220901</enddate><creator>Conzentino, Marcelo S.</creator><creator>Gonçalves, Ana C. A.</creator><creator>Paula, Nigella M.</creator><creator>Rego, Fabiane G. M.</creator><creator>Zanette, Dalila L.</creator><creator>Aoki, Mateus N.</creator><creator>Nardin, Jeanine M.</creator><creator>Huergo, Luciano Fernandes</creator><general>Springer International Publishing</general><general>Springer Nature B.V</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7U9</scope><scope>C1K</scope><scope>H94</scope><scope>K9.</scope><scope>M7N</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20220901</creationdate><title>A magnetic bead immunoassay to detect high affinity human IgG reactive to SARS-CoV-2 Spike S1 RBD produced in Escherichia coli</title><author>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</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c474t-e428d5a0c21484e24e5238f8f4484bd0ff89b424c5259c93bd2540f7c40106063</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Affinity</topic><topic>Affinity chromatography</topic><topic>Antibodies</topic><topic>Antigens</topic><topic>Beads</topic><topic>Binding</topic><topic>Biomedical and Life Sciences</topic><topic>Cell lines</topic><topic>Clinical Microbiology - Research Paper</topic><topic>COVID-19</topic><topic>Domains</topic><topic>E coli</topic><topic>Escherichia coli</topic><topic>Food Microbiology</topic><topic>Human populations</topic><topic>Immunization</topic><topic>Immunoassay</topic><topic>Immunoglobulin G</topic><topic>Immunology</topic><topic>Life Sciences</topic><topic>Low cost</topic><topic>Medical Microbiology</topic><topic>Microbial Ecology</topic><topic>Microbial Genetics and Genomics</topic><topic>Microbiology</topic><topic>Mycology</topic><topic>Pandemics</topic><topic>Public health</topic><topic>Receptors</topic><topic>Sensitivity</topic><topic>Seroconversion</topic><topic>Severe acute respiratory syndrome</topic><topic>Severe acute respiratory syndrome coronavirus 2</topic><topic>Vaccination</topic><topic>Vaccines</topic><topic>Viral diseases</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Conzentino, Marcelo S.</creatorcontrib><creatorcontrib>Gonçalves, Ana C. A.</creatorcontrib><creatorcontrib>Paula, Nigella M.</creatorcontrib><creatorcontrib>Rego, Fabiane G. M.</creatorcontrib><creatorcontrib>Zanette, Dalila L.</creatorcontrib><creatorcontrib>Aoki, Mateus N.</creatorcontrib><creatorcontrib>Nardin, Jeanine M.</creatorcontrib><creatorcontrib>Huergo, Luciano Fernandes</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Virology and AIDS Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Brazilian journal of microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Conzentino, Marcelo S.</au><au>Gonçalves, Ana C. A.</au><au>Paula, Nigella M.</au><au>Rego, Fabiane G. M.</au><au>Zanette, Dalila L.</au><au>Aoki, Mateus N.</au><au>Nardin, Jeanine M.</au><au>Huergo, Luciano Fernandes</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A magnetic bead immunoassay to detect high affinity human IgG reactive to SARS-CoV-2 Spike S1 RBD produced in Escherichia coli</atitle><jtitle>Brazilian journal of microbiology</jtitle><stitle>Braz J Microbiol</stitle><addtitle>Braz J Microbiol</addtitle><date>2022-09-01</date><risdate>2022</risdate><volume>53</volume><issue>3</issue><spage>1263</spage><epage>1269</epage><pages>1263-1269</pages><issn>1517-8382</issn><eissn>1678-4405</eissn><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
2+
magnetic bead immunoassay to detect human IgG reactive to Spike S1 RBD Receptor Binding Domain produced in
Escherichia coli.
A 6xHis-tagged Spike S1 RBD was expressed in
E. coli
and purified by affinity chromatography. The protein was mobilized on the surface of Ni
2+
magnetic beads and used to investigate the presence of reactive IgG in the serum obtained from pre-pandemic and COVID-19 confirmed cases. The method was validated with a cohort of 290 samples and an area under the receiver operating characteristic curve of 0.94 was obtained. The method was operated with > 82% sensitivity at 98% specificity and was also able to track human IgG raised in response to vaccination with Comirnaty at > 85% sensitivity. The IgG signal obtained with the described method was well-correlated with the signal obtained when pre fusion Spike produced in HEK cell lines was used as antigen. This novel low-cost and high throughput immunoassay may act as an important tool to investigate protecting IgG antibodies against SARS-CoV-2 in the human population.</abstract><cop>Cham</cop><pub>Springer International Publishing</pub><pmid>35426068</pmid><doi>10.1007/s42770-022-00753-x</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1517-8382 |
| ispartof | Brazilian journal of microbiology, 2022-09, Vol.53 (3), p.1263-1269 |
| issn | 1517-8382 1678-4405 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_9009495 |
| source | SpringerLink (Online service); PubMed Central; Alma/SFX Local Collection |
| subjects | Affinity Affinity chromatography Antibodies Antigens Beads Binding Biomedical and Life Sciences Cell lines Clinical Microbiology - Research Paper COVID-19 Domains E coli Escherichia coli Food Microbiology Human populations Immunization Immunoassay Immunoglobulin G Immunology Life Sciences Low cost Medical Microbiology Microbial Ecology Microbial Genetics and Genomics Microbiology Mycology Pandemics Public health Receptors Sensitivity Seroconversion Severe acute respiratory syndrome Severe acute respiratory syndrome coronavirus 2 Vaccination Vaccines Viral diseases |
| title | A magnetic bead immunoassay to detect high affinity human IgG reactive to SARS-CoV-2 Spike S1 RBD produced in Escherichia coli |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-14T18%3A53%3A58IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=A%20magnetic%20bead%20immunoassay%20to%20detect%20high%20affinity%20human%20IgG%20reactive%20to%20SARS-CoV-2%20Spike%20S1%20RBD%20produced%20in%20Escherichia%20coli&rft.jtitle=Brazilian%20journal%20of%20microbiology&rft.au=Conzentino,%20Marcelo%20S.&rft.date=2022-09-01&rft.volume=53&rft.issue=3&rft.spage=1263&rft.epage=1269&rft.pages=1263-1269&rft.issn=1517-8382&rft.eissn=1678-4405&rft_id=info:doi/10.1007/s42770-022-00753-x&rft_dat=%3Cproquest_pubme%3E2651693605%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2708764053&rft_id=info:pmid/35426068&rfr_iscdi=true |