An anti-phospholipase A2 receptor quantitative immunoassay and epitope analysis in membranous nephropathy reveals different antigenic domains of the receptor
The phospholipase A2 receptor (PLA2R) was recently discovered as a target autoantigen in patients with idiopathic membranous nephropathy (IMN). Published evidence suggests that the autoantibodies directed towards a conformation dependent epitope are currently effectively detected by a cell based ass...
Gespeichert in:
| Veröffentlicht in: | PloS one 2013-04, Vol.8 (4), p.e61669-e61669 |
|---|---|
| Hauptverfasser: | , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | e61669 |
|---|---|
| container_issue | 4 |
| container_start_page | e61669 |
| container_title | PloS one |
| container_volume | 8 |
| creator | Behnert, Astrid Fritzler, Marvin J Teng, Beina Zhang, Meifeng Bollig, Frank Haller, Hermann Skoberne, Andrej Mahler, Michael Schiffer, Mario |
| description | The phospholipase A2 receptor (PLA2R) was recently discovered as a target autoantigen in patients with idiopathic membranous nephropathy (IMN). Published evidence suggests that the autoantibodies directed towards a conformation dependent epitope are currently effectively detected by a cell based assay (CBA) utilizing indirect immunofluorescence (IIF) on tissue culture cells transfected with the PLA2R cDNA. Limitations of such IIF-CBA assays include observer dependent subjective evaluation of semi-quantitative test results and the protocols are not amenable to high throughput diagnostic testing. We developed a quantitative, observer independent, high throughput capture immunoassay for detecting PLA2R autoantibodies on an addressable laser bead immunoassay (ALBIA) platform. Since reactive domains of PLA2R (i.e. epitopes) could be used to improve diagnostic tests by using small peptides in various high throughput diagnostic platforms, we identified PLA2R epitopes that bound autoantibodies of IMN patients. These studies confirmed that inter-molecular epitope spreading occurs in IMN but use of the cognate synthetic peptides in immunoassays was unable to conclusively distinguish between IMN patients and normal controls. However, combinations of these peptides were able to effectively absorb anti-PLA2R reactivity in IIF-CBA and an immunoassay that employed a lysate derived from HEK cells tranfected with and overexpressing PLA2R. While we provide evidence of intermolecular epitope spreading, our data indicates that in addition to conformational epitopes, human anti-PLA2R reactivity in a commercially available CBA and an addressable laser bead immunoassay is significantly absorbed by peptides representing epitopes of PLA2R. |
| doi_str_mv | 10.1371/journal.pone.0061669 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_plos_</sourceid><recordid>TN_cdi_plos_journals_1347295845</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><doaj_id>oai_doaj_org_article_20e17bc829c645a59d5b23f769528b16</doaj_id><sourcerecordid>1348500606</sourcerecordid><originalsourceid>FETCH-LOGICAL-c5079-5f4fbeb191fa0621d52014641162b4adfe0523f8e938918577ac1c3c6de4430e3</originalsourceid><addsrcrecordid>eNptUstq3DAUNaWlSdP-QWkF3XTjqR6WbG0KQ-gjEOimXQtZvh5rsCVHsgfmY_KvlTPOkJQuhIR0HvdenSx7T_CGsJJ82fs5ON1vRu9gg7EgQsgX2SWRjOaCYvbyyfkiexPjHmPOKiFeZxeUCVZWpbzM7rcOaTfZfOx8TKu3o46AthQFMDBOPqC7eQFMerIHQHYYZud1jPqYeA2C0U5-hHTW_THaiKxDAwx10M7PETkYu-BHPXXHJHgA3UfU2LaFAG56MN6BswY1ftDWReRbNHVw9n6bvWoTBd6t-1X25_u339c_89tfP26ut7e54biUOW-LtoaaSNJqLChpOMWkEAUhgtaFblrAnLK2AskqSSpeltoQw4xooCgYBnaVfTzpjr2Pap1sVIQVJZW8KnhC3JwQjdd7NQY76HBUXlv1cOHDTukwWdODohhIWZuKSiMKrrlseJ3cSyE5rWoiktbX1W2uB2hMGkXQ_TPR5y_OdmrnDyr9mqR8KebzKhD83QxxUoONBvpeO0hTX-queIoEXrw-_QP9f3fFCWWCjzFAey6GYLWk7ZGllrSpNW2J9uFpI2fSY7zYXx0Z1rA</addsrcrecordid><sourcetype>Open Website</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1347295845</pqid></control><display><type>article</type><title>An anti-phospholipase A2 receptor quantitative immunoassay and epitope analysis in membranous nephropathy reveals different antigenic domains of the receptor</title><source>MEDLINE</source><source>DOAJ Directory of Open Access Journals</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>Public Library of Science (PLoS) Journals Open Access</source><source>PubMed Central</source><source>Free Full-Text Journals in Chemistry</source><creator>Behnert, Astrid ; Fritzler, Marvin J ; Teng, Beina ; Zhang, Meifeng ; Bollig, Frank ; Haller, Hermann ; Skoberne, Andrej ; Mahler, Michael ; Schiffer, Mario</creator><contributor>Pizzo, Salvatore V.</contributor><creatorcontrib>Behnert, Astrid ; Fritzler, Marvin J ; Teng, Beina ; Zhang, Meifeng ; Bollig, Frank ; Haller, Hermann ; Skoberne, Andrej ; Mahler, Michael ; Schiffer, Mario ; Pizzo, Salvatore V.</creatorcontrib><description>The phospholipase A2 receptor (PLA2R) was recently discovered as a target autoantigen in patients with idiopathic membranous nephropathy (IMN). Published evidence suggests that the autoantibodies directed towards a conformation dependent epitope are currently effectively detected by a cell based assay (CBA) utilizing indirect immunofluorescence (IIF) on tissue culture cells transfected with the PLA2R cDNA. Limitations of such IIF-CBA assays include observer dependent subjective evaluation of semi-quantitative test results and the protocols are not amenable to high throughput diagnostic testing. We developed a quantitative, observer independent, high throughput capture immunoassay for detecting PLA2R autoantibodies on an addressable laser bead immunoassay (ALBIA) platform. Since reactive domains of PLA2R (i.e. epitopes) could be used to improve diagnostic tests by using small peptides in various high throughput diagnostic platforms, we identified PLA2R epitopes that bound autoantibodies of IMN patients. These studies confirmed that inter-molecular epitope spreading occurs in IMN but use of the cognate synthetic peptides in immunoassays was unable to conclusively distinguish between IMN patients and normal controls. However, combinations of these peptides were able to effectively absorb anti-PLA2R reactivity in IIF-CBA and an immunoassay that employed a lysate derived from HEK cells tranfected with and overexpressing PLA2R. While we provide evidence of intermolecular epitope spreading, our data indicates that in addition to conformational epitopes, human anti-PLA2R reactivity in a commercially available CBA and an addressable laser bead immunoassay is significantly absorbed by peptides representing epitopes of PLA2R.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0061669</identifier><identifier>PMID: 23637879</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Amino Acid Sequence ; Antigens ; Arthritis ; Autoantibodies ; Autoantibodies - analysis ; Autoantibodies - immunology ; Autoantigens - chemistry ; Autoantigens - immunology ; Biology ; Cell culture ; Conformation ; Diagnostic systems ; Disease ; Epitope Mapping - methods ; Epitopes ; Glomerulonephritis, Membranous - diagnosis ; HEK293 Cells ; Humans ; Immunoassay ; Immunoassay - methods ; Immunoassays ; Immunofluorescence ; Immunoglobulins ; Lasers ; Medical schools ; Medicine ; Membranous nephropathy ; Molecular Sequence Data ; Nephrology ; Nephropathy ; Patients ; Peptide Fragments - chemistry ; Peptide Fragments - immunology ; Peptides ; Phospholipase ; Phospholipase A2 ; Protein Structure, Tertiary ; Receptors, Phospholipase A2 - chemistry ; Receptors, Phospholipase A2 - immunology ; Rodents ; Spreading ; Synthetic peptides ; Tissue culture</subject><ispartof>PloS one, 2013-04, Vol.8 (4), p.e61669-e61669</ispartof><rights>2013 Behnert et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2013 Behnert et al 2013 Behnert et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5079-5f4fbeb191fa0621d52014641162b4adfe0523f8e938918577ac1c3c6de4430e3</citedby><cites>FETCH-LOGICAL-c5079-5f4fbeb191fa0621d52014641162b4adfe0523f8e938918577ac1c3c6de4430e3</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/PMC3639255/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3639255/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2102,2928,23866,27924,27925,53791,53793,79600,79601</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23637879$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Pizzo, Salvatore V.</contributor><creatorcontrib>Behnert, Astrid</creatorcontrib><creatorcontrib>Fritzler, Marvin J</creatorcontrib><creatorcontrib>Teng, Beina</creatorcontrib><creatorcontrib>Zhang, Meifeng</creatorcontrib><creatorcontrib>Bollig, Frank</creatorcontrib><creatorcontrib>Haller, Hermann</creatorcontrib><creatorcontrib>Skoberne, Andrej</creatorcontrib><creatorcontrib>Mahler, Michael</creatorcontrib><creatorcontrib>Schiffer, Mario</creatorcontrib><title>An anti-phospholipase A2 receptor quantitative immunoassay and epitope analysis in membranous nephropathy reveals different antigenic domains of the receptor</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>The phospholipase A2 receptor (PLA2R) was recently discovered as a target autoantigen in patients with idiopathic membranous nephropathy (IMN). Published evidence suggests that the autoantibodies directed towards a conformation dependent epitope are currently effectively detected by a cell based assay (CBA) utilizing indirect immunofluorescence (IIF) on tissue culture cells transfected with the PLA2R cDNA. Limitations of such IIF-CBA assays include observer dependent subjective evaluation of semi-quantitative test results and the protocols are not amenable to high throughput diagnostic testing. We developed a quantitative, observer independent, high throughput capture immunoassay for detecting PLA2R autoantibodies on an addressable laser bead immunoassay (ALBIA) platform. Since reactive domains of PLA2R (i.e. epitopes) could be used to improve diagnostic tests by using small peptides in various high throughput diagnostic platforms, we identified PLA2R epitopes that bound autoantibodies of IMN patients. These studies confirmed that inter-molecular epitope spreading occurs in IMN but use of the cognate synthetic peptides in immunoassays was unable to conclusively distinguish between IMN patients and normal controls. However, combinations of these peptides were able to effectively absorb anti-PLA2R reactivity in IIF-CBA and an immunoassay that employed a lysate derived from HEK cells tranfected with and overexpressing PLA2R. While we provide evidence of intermolecular epitope spreading, our data indicates that in addition to conformational epitopes, human anti-PLA2R reactivity in a commercially available CBA and an addressable laser bead immunoassay is significantly absorbed by peptides representing epitopes of PLA2R.</description><subject>Amino Acid Sequence</subject><subject>Antigens</subject><subject>Arthritis</subject><subject>Autoantibodies</subject><subject>Autoantibodies - analysis</subject><subject>Autoantibodies - immunology</subject><subject>Autoantigens - chemistry</subject><subject>Autoantigens - immunology</subject><subject>Biology</subject><subject>Cell culture</subject><subject>Conformation</subject><subject>Diagnostic systems</subject><subject>Disease</subject><subject>Epitope Mapping - methods</subject><subject>Epitopes</subject><subject>Glomerulonephritis, Membranous - diagnosis</subject><subject>HEK293 Cells</subject><subject>Humans</subject><subject>Immunoassay</subject><subject>Immunoassay - methods</subject><subject>Immunoassays</subject><subject>Immunofluorescence</subject><subject>Immunoglobulins</subject><subject>Lasers</subject><subject>Medical schools</subject><subject>Medicine</subject><subject>Membranous nephropathy</subject><subject>Molecular Sequence Data</subject><subject>Nephrology</subject><subject>Nephropathy</subject><subject>Patients</subject><subject>Peptide Fragments - chemistry</subject><subject>Peptide Fragments - immunology</subject><subject>Peptides</subject><subject>Phospholipase</subject><subject>Phospholipase A2</subject><subject>Protein Structure, Tertiary</subject><subject>Receptors, Phospholipase A2 - chemistry</subject><subject>Receptors, Phospholipase A2 - immunology</subject><subject>Rodents</subject><subject>Spreading</subject><subject>Synthetic peptides</subject><subject>Tissue culture</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>DOA</sourceid><recordid>eNptUstq3DAUNaWlSdP-QWkF3XTjqR6WbG0KQ-gjEOimXQtZvh5rsCVHsgfmY_KvlTPOkJQuhIR0HvdenSx7T_CGsJJ82fs5ON1vRu9gg7EgQsgX2SWRjOaCYvbyyfkiexPjHmPOKiFeZxeUCVZWpbzM7rcOaTfZfOx8TKu3o46AthQFMDBOPqC7eQFMerIHQHYYZud1jPqYeA2C0U5-hHTW_THaiKxDAwx10M7PETkYu-BHPXXHJHgA3UfU2LaFAG56MN6BswY1ftDWReRbNHVw9n6bvWoTBd6t-1X25_u339c_89tfP26ut7e54biUOW-LtoaaSNJqLChpOMWkEAUhgtaFblrAnLK2AskqSSpeltoQw4xooCgYBnaVfTzpjr2Pap1sVIQVJZW8KnhC3JwQjdd7NQY76HBUXlv1cOHDTukwWdODohhIWZuKSiMKrrlseJ3cSyE5rWoiktbX1W2uB2hMGkXQ_TPR5y_OdmrnDyr9mqR8KebzKhD83QxxUoONBvpeO0hTX-queIoEXrw-_QP9f3fFCWWCjzFAey6GYLWk7ZGllrSpNW2J9uFpI2fSY7zYXx0Z1rA</recordid><startdate>20130429</startdate><enddate>20130429</enddate><creator>Behnert, Astrid</creator><creator>Fritzler, Marvin J</creator><creator>Teng, Beina</creator><creator>Zhang, Meifeng</creator><creator>Bollig, Frank</creator><creator>Haller, Hermann</creator><creator>Skoberne, Andrej</creator><creator>Mahler, Michael</creator><creator>Schiffer, Mario</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20130429</creationdate><title>An anti-phospholipase A2 receptor quantitative immunoassay and epitope analysis in membranous nephropathy reveals different antigenic domains of the receptor</title><author>Behnert, Astrid ; Fritzler, Marvin J ; Teng, Beina ; Zhang, Meifeng ; Bollig, Frank ; Haller, Hermann ; Skoberne, Andrej ; Mahler, Michael ; Schiffer, Mario</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5079-5f4fbeb191fa0621d52014641162b4adfe0523f8e938918577ac1c3c6de4430e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Amino Acid Sequence</topic><topic>Antigens</topic><topic>Arthritis</topic><topic>Autoantibodies</topic><topic>Autoantibodies - analysis</topic><topic>Autoantibodies - immunology</topic><topic>Autoantigens - chemistry</topic><topic>Autoantigens - immunology</topic><topic>Biology</topic><topic>Cell culture</topic><topic>Conformation</topic><topic>Diagnostic systems</topic><topic>Disease</topic><topic>Epitope Mapping - methods</topic><topic>Epitopes</topic><topic>Glomerulonephritis, Membranous - diagnosis</topic><topic>HEK293 Cells</topic><topic>Humans</topic><topic>Immunoassay</topic><topic>Immunoassay - methods</topic><topic>Immunoassays</topic><topic>Immunofluorescence</topic><topic>Immunoglobulins</topic><topic>Lasers</topic><topic>Medical schools</topic><topic>Medicine</topic><topic>Membranous nephropathy</topic><topic>Molecular Sequence Data</topic><topic>Nephrology</topic><topic>Nephropathy</topic><topic>Patients</topic><topic>Peptide Fragments - chemistry</topic><topic>Peptide Fragments - immunology</topic><topic>Peptides</topic><topic>Phospholipase</topic><topic>Phospholipase A2</topic><topic>Protein Structure, Tertiary</topic><topic>Receptors, Phospholipase A2 - chemistry</topic><topic>Receptors, Phospholipase A2 - immunology</topic><topic>Rodents</topic><topic>Spreading</topic><topic>Synthetic peptides</topic><topic>Tissue culture</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Behnert, Astrid</creatorcontrib><creatorcontrib>Fritzler, Marvin J</creatorcontrib><creatorcontrib>Teng, Beina</creatorcontrib><creatorcontrib>Zhang, Meifeng</creatorcontrib><creatorcontrib>Bollig, Frank</creatorcontrib><creatorcontrib>Haller, Hermann</creatorcontrib><creatorcontrib>Skoberne, Andrej</creatorcontrib><creatorcontrib>Mahler, Michael</creatorcontrib><creatorcontrib>Schiffer, Mario</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - Academic</collection><collection>ProQuest Engineering Collection</collection><collection>ProQuest Biological Science Collection</collection><collection>Agricultural Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Engineering Database</collection><collection>Nursing & Allied Health Premium</collection><collection>Advanced Technologies & Aerospace Database</collection><collection>ProQuest Advanced Technologies & Aerospace Collection</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Environmental Science Database</collection><collection>Materials Science Collection</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>Engineering Collection</collection><collection>Environmental Science Collection</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Behnert, Astrid</au><au>Fritzler, Marvin J</au><au>Teng, Beina</au><au>Zhang, Meifeng</au><au>Bollig, Frank</au><au>Haller, Hermann</au><au>Skoberne, Andrej</au><au>Mahler, Michael</au><au>Schiffer, Mario</au><au>Pizzo, Salvatore V.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>An anti-phospholipase A2 receptor quantitative immunoassay and epitope analysis in membranous nephropathy reveals different antigenic domains of the receptor</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2013-04-29</date><risdate>2013</risdate><volume>8</volume><issue>4</issue><spage>e61669</spage><epage>e61669</epage><pages>e61669-e61669</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>The phospholipase A2 receptor (PLA2R) was recently discovered as a target autoantigen in patients with idiopathic membranous nephropathy (IMN). Published evidence suggests that the autoantibodies directed towards a conformation dependent epitope are currently effectively detected by a cell based assay (CBA) utilizing indirect immunofluorescence (IIF) on tissue culture cells transfected with the PLA2R cDNA. Limitations of such IIF-CBA assays include observer dependent subjective evaluation of semi-quantitative test results and the protocols are not amenable to high throughput diagnostic testing. We developed a quantitative, observer independent, high throughput capture immunoassay for detecting PLA2R autoantibodies on an addressable laser bead immunoassay (ALBIA) platform. Since reactive domains of PLA2R (i.e. epitopes) could be used to improve diagnostic tests by using small peptides in various high throughput diagnostic platforms, we identified PLA2R epitopes that bound autoantibodies of IMN patients. These studies confirmed that inter-molecular epitope spreading occurs in IMN but use of the cognate synthetic peptides in immunoassays was unable to conclusively distinguish between IMN patients and normal controls. However, combinations of these peptides were able to effectively absorb anti-PLA2R reactivity in IIF-CBA and an immunoassay that employed a lysate derived from HEK cells tranfected with and overexpressing PLA2R. While we provide evidence of intermolecular epitope spreading, our data indicates that in addition to conformational epitopes, human anti-PLA2R reactivity in a commercially available CBA and an addressable laser bead immunoassay is significantly absorbed by peptides representing epitopes of PLA2R.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>23637879</pmid><doi>10.1371/journal.pone.0061669</doi><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2013-04, Vol.8 (4), p.e61669-e61669 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1347295845 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Public Library of Science (PLoS) Journals Open Access; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Amino Acid Sequence Antigens Arthritis Autoantibodies Autoantibodies - analysis Autoantibodies - immunology Autoantigens - chemistry Autoantigens - immunology Biology Cell culture Conformation Diagnostic systems Disease Epitope Mapping - methods Epitopes Glomerulonephritis, Membranous - diagnosis HEK293 Cells Humans Immunoassay Immunoassay - methods Immunoassays Immunofluorescence Immunoglobulins Lasers Medical schools Medicine Membranous nephropathy Molecular Sequence Data Nephrology Nephropathy Patients Peptide Fragments - chemistry Peptide Fragments - immunology Peptides Phospholipase Phospholipase A2 Protein Structure, Tertiary Receptors, Phospholipase A2 - chemistry Receptors, Phospholipase A2 - immunology Rodents Spreading Synthetic peptides Tissue culture |
| title | An anti-phospholipase A2 receptor quantitative immunoassay and epitope analysis in membranous nephropathy reveals different antigenic domains of the receptor |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-04T15%3A03%3A36IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=An%20anti-phospholipase%20A2%20receptor%20quantitative%20immunoassay%20and%20epitope%20analysis%20in%20membranous%20nephropathy%20reveals%20different%20antigenic%20domains%20of%20the%20receptor&rft.jtitle=PloS%20one&rft.au=Behnert,%20Astrid&rft.date=2013-04-29&rft.volume=8&rft.issue=4&rft.spage=e61669&rft.epage=e61669&rft.pages=e61669-e61669&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0061669&rft_dat=%3Cproquest_plos_%3E1348500606%3C/proquest_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1347295845&rft_id=info:pmid/23637879&rft_doaj_id=oai_doaj_org_article_20e17bc829c645a59d5b23f769528b16&rfr_iscdi=true |