Recombinant protein to analyze autoantibodies to proteinase 3 in systemic vasculitis
The presence of antineutrophil cytoplasmic autoantibodies with specificity for proteinase 3 (PR3-ANCA) usually is detected by enzyme-linked immunosorbent assay (ELISA) with purified PR3 as a substrate. We studied the technical performance of direct and capture ELISA using a recombinant proteolytical...
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| Veröffentlicht in: | American journal of clinical pathology 2003-10, Vol.120 (4), p.586-595 |
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| creator | RAROK, Agnieszka A HUITEMA, Minke G VAN DER LEIJ, Marcel J VAN DER GELD, Ymke M BERTHOLD, Heike SCHMITT, Jacky STEGEMAN, Coen A LIMBURG, Pieter C KALLENBERG, Cees G. M |
| description | The presence of antineutrophil cytoplasmic autoantibodies with specificity for proteinase 3 (PR3-ANCA) usually is detected by enzyme-linked immunosorbent assay (ELISA) with purified PR3 as a substrate. We studied the technical performance of direct and capture ELISA using a recombinant proteolytically inactive form of PR3 produced in the baculovirus expression system for the detection of PR3-ANCA in 114 patients with systemic vasculitis at diagnosis. We found that ELISA using recombinant PR3 produced in insect cells is a promising alternative for ELISA with native PR3. We found a correlation between tests using recombinant or native PR3, as well as correlation of the ELISA results with ANCA titers measured by the indirect immunofluorescence technique. However, the specificity for ANCA-associated vasculitis of ELISA with recombinant PR3 was lower than ELISA using native PR3. Compared with the direct assay, capture ELISA is a more sensitive method for PR3-ANCA detection, with both native and recombinant PR3, and its results depend on the monoclonal antibody used to capture the antigen. |
| doi_str_mv | 10.1309/YTU2FUHBEXJLWMLD |
| format | Article |
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M</creator><creatorcontrib>RAROK, Agnieszka A ; HUITEMA, Minke G ; VAN DER LEIJ, Marcel J ; VAN DER GELD, Ymke M ; BERTHOLD, Heike ; SCHMITT, Jacky ; STEGEMAN, Coen A ; LIMBURG, Pieter C ; KALLENBERG, Cees G. M</creatorcontrib><description>The presence of antineutrophil cytoplasmic autoantibodies with specificity for proteinase 3 (PR3-ANCA) usually is detected by enzyme-linked immunosorbent assay (ELISA) with purified PR3 as a substrate. We studied the technical performance of direct and capture ELISA using a recombinant proteolytically inactive form of PR3 produced in the baculovirus expression system for the detection of PR3-ANCA in 114 patients with systemic vasculitis at diagnosis. We found that ELISA using recombinant PR3 produced in insect cells is a promising alternative for ELISA with native PR3. We found a correlation between tests using recombinant or native PR3, as well as correlation of the ELISA results with ANCA titers measured by the indirect immunofluorescence technique. However, the specificity for ANCA-associated vasculitis of ELISA with recombinant PR3 was lower than ELISA using native PR3. Compared with the direct assay, capture ELISA is a more sensitive method for PR3-ANCA detection, with both native and recombinant PR3, and its results depend on the monoclonal antibody used to capture the antigen.</description><identifier>ISSN: 0002-9173</identifier><identifier>EISSN: 1943-7722</identifier><identifier>DOI: 10.1309/YTU2FUHBEXJLWMLD</identifier><identifier>PMID: 14560570</identifier><identifier>CODEN: AJCPAI</identifier><language>eng</language><publisher>Chicago, IL: American Society of Clinical Pathologists</publisher><subject>Animals ; Antibodies, Antineutrophil Cytoplasmic - immunology ; Antibodies, Monoclonal ; Baculovirus ; Biological and medical sciences ; Cells, Cultured ; Enzyme-Linked Immunosorbent Assay - methods ; Female ; Fluorescent Antibody Technique, Indirect ; Genetic Vectors ; Humans ; Male ; Medical sciences ; Myeloblastin ; Recombinant Proteins - immunology ; Recombinant Proteins - metabolism ; Sarcoidosis. Granulomatous diseases of unproved etiology. 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Vasculitis ; Sensitivity and Specificity ; Serine Endopeptidases - immunology ; Serine Endopeptidases - metabolism ; Spodoptera - genetics ; Spodoptera - metabolism ; Vasculitis - diagnosis ; Vasculitis - enzymology ; Vasculitis - immunology ; Viral Proteins - immunology ; Viral Proteins - metabolism</subject><ispartof>American journal of clinical pathology, 2003-10, Vol.120 (4), p.586-595</ispartof><rights>2004 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c398t-3c130807a2207d4043a4e7cb3f6bb44e46ee6c10fe4c8ae851f6aaaf9fc0d6a23</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27922,27923</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=15178015$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/14560570$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>RAROK, Agnieszka A</creatorcontrib><creatorcontrib>HUITEMA, Minke G</creatorcontrib><creatorcontrib>VAN DER LEIJ, Marcel J</creatorcontrib><creatorcontrib>VAN DER GELD, Ymke M</creatorcontrib><creatorcontrib>BERTHOLD, Heike</creatorcontrib><creatorcontrib>SCHMITT, Jacky</creatorcontrib><creatorcontrib>STEGEMAN, Coen A</creatorcontrib><creatorcontrib>LIMBURG, Pieter C</creatorcontrib><creatorcontrib>KALLENBERG, Cees G. M</creatorcontrib><title>Recombinant protein to analyze autoantibodies to proteinase 3 in systemic vasculitis</title><title>American journal of clinical pathology</title><addtitle>Am J Clin Pathol</addtitle><description>The presence of antineutrophil cytoplasmic autoantibodies with specificity for proteinase 3 (PR3-ANCA) usually is detected by enzyme-linked immunosorbent assay (ELISA) with purified PR3 as a substrate. We studied the technical performance of direct and capture ELISA using a recombinant proteolytically inactive form of PR3 produced in the baculovirus expression system for the detection of PR3-ANCA in 114 patients with systemic vasculitis at diagnosis. We found that ELISA using recombinant PR3 produced in insect cells is a promising alternative for ELISA with native PR3. We found a correlation between tests using recombinant or native PR3, as well as correlation of the ELISA results with ANCA titers measured by the indirect immunofluorescence technique. However, the specificity for ANCA-associated vasculitis of ELISA with recombinant PR3 was lower than ELISA using native PR3. Compared with the direct assay, capture ELISA is a more sensitive method for PR3-ANCA detection, with both native and recombinant PR3, and its results depend on the monoclonal antibody used to capture the antigen.</description><subject>Animals</subject><subject>Antibodies, Antineutrophil Cytoplasmic - immunology</subject><subject>Antibodies, Monoclonal</subject><subject>Baculovirus</subject><subject>Biological and medical sciences</subject><subject>Cells, Cultured</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>Female</subject><subject>Fluorescent Antibody Technique, Indirect</subject><subject>Genetic Vectors</subject><subject>Humans</subject><subject>Male</subject><subject>Medical sciences</subject><subject>Myeloblastin</subject><subject>Recombinant Proteins - immunology</subject><subject>Recombinant Proteins - metabolism</subject><subject>Sarcoidosis. Granulomatous diseases of unproved etiology. Connective tissue diseases. Elastic tissue diseases. Vasculitis</subject><subject>Sensitivity and Specificity</subject><subject>Serine Endopeptidases - immunology</subject><subject>Serine Endopeptidases - metabolism</subject><subject>Spodoptera - genetics</subject><subject>Spodoptera - metabolism</subject><subject>Vasculitis - diagnosis</subject><subject>Vasculitis - enzymology</subject><subject>Vasculitis - immunology</subject><subject>Viral Proteins - immunology</subject><subject>Viral Proteins - metabolism</subject><issn>0002-9173</issn><issn>1943-7722</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1Lw0AQhhdRbK3ePUkueovOfiSbHLW2VqkI0qKewmQ7gZV81Gwi1F9vSgMFL57mMM_7MvMwds7hmkuIbz4WSzFdzu4m70_zt-f5_QEb8lhJX2shDtkQAIQfcy0H7MS5TwAuIlDHbMBVEEKgYcgWr2SqIrUllo23rquGbOk1lYcl5psf8rBtqm5l02plyW03PYSOPOl1sNu4hgprvG90ps1tY90pO8owd3TWzxFbTieL8cyfvzw8jm_nvpFx1PjSdE9EoFEI0CsFSqIibVKZhWmqFKmQKDQcMlImQooCnoWImMWZgVWIQo7Y1a63u-mrJdckhXWG8hxLqlqXaC50qEP-L8jjWEAURB0IO9DUlXM1Zcm6tgXWm4RDslWe_FXeRS767jYtaLUP9I474LIHOkGYZzWWxro9F3AdAQ_kL5yfjII</recordid><startdate>20031001</startdate><enddate>20031001</enddate><creator>RAROK, Agnieszka A</creator><creator>HUITEMA, Minke G</creator><creator>VAN DER LEIJ, Marcel J</creator><creator>VAN DER GELD, Ymke M</creator><creator>BERTHOLD, Heike</creator><creator>SCHMITT, Jacky</creator><creator>STEGEMAN, Coen A</creator><creator>LIMBURG, Pieter C</creator><creator>KALLENBERG, Cees G. 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M</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Recombinant protein to analyze autoantibodies to proteinase 3 in systemic vasculitis</atitle><jtitle>American journal of clinical pathology</jtitle><addtitle>Am J Clin Pathol</addtitle><date>2003-10-01</date><risdate>2003</risdate><volume>120</volume><issue>4</issue><spage>586</spage><epage>595</epage><pages>586-595</pages><issn>0002-9173</issn><eissn>1943-7722</eissn><coden>AJCPAI</coden><abstract>The presence of antineutrophil cytoplasmic autoantibodies with specificity for proteinase 3 (PR3-ANCA) usually is detected by enzyme-linked immunosorbent assay (ELISA) with purified PR3 as a substrate. We studied the technical performance of direct and capture ELISA using a recombinant proteolytically inactive form of PR3 produced in the baculovirus expression system for the detection of PR3-ANCA in 114 patients with systemic vasculitis at diagnosis. 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| source | MEDLINE; Oxford University Press Journals All Titles (1996-Current); EZB-FREE-00999 freely available EZB journals |
| subjects | Animals Antibodies, Antineutrophil Cytoplasmic - immunology Antibodies, Monoclonal Baculovirus Biological and medical sciences Cells, Cultured Enzyme-Linked Immunosorbent Assay - methods Female Fluorescent Antibody Technique, Indirect Genetic Vectors Humans Male Medical sciences Myeloblastin Recombinant Proteins - immunology Recombinant Proteins - metabolism Sarcoidosis. Granulomatous diseases of unproved etiology. Connective tissue diseases. Elastic tissue diseases. Vasculitis Sensitivity and Specificity Serine Endopeptidases - immunology Serine Endopeptidases - metabolism Spodoptera - genetics Spodoptera - metabolism Vasculitis - diagnosis Vasculitis - enzymology Vasculitis - immunology Viral Proteins - immunology Viral Proteins - metabolism |
| title | Recombinant protein to analyze autoantibodies to proteinase 3 in systemic vasculitis |
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