Detection of anti‐topoisomerase I antibodies using a full length human topoisomerase I recombinant protein purified from a baculovirus expression system
SUMMARY Topoisomerase I (topo I) is a major systemic sclerosis (SSc)‐associated autoantigen. A cDNA construct encoding full length human topo I in a recombinant baculovirus transfer vector was used to infect insect cells in culture from which recombinant protein was purified. An ELISA using recombin...
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| Veröffentlicht in: | Clinical and experimental immunology 1995-05, Vol.100 (2), p.214-218 |
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| container_title | Clinical and experimental immunology |
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| creator | WHYTE, J. EARNSHAW, W. C. CHAMPOUX, J. J. PARKER, L. H. STEWART, L. HALL, N. D. MCHUGH, N. J. |
| description | SUMMARY
Topoisomerase I (topo I) is a major systemic sclerosis (SSc)‐associated autoantigen. A cDNA construct encoding full length human topo I in a recombinant baculovirus transfer vector was used to infect insect cells in culture from which recombinant protein was purified. An ELISA using recombinant protein was evaluated in 340 sera including sera from 134 patients with SSc, of whom 33 had anti‐topo I antibodies detected by immunodiffusion. A high yield of pure topo I of expected molecular mass and catalytic activity was obtained. The recombinant topo I ELISA was 92% sensitive and 98% specific in detecting anti‐topo I antibodies which were present almost exclusively in patients with SSc. Therefore, the potential advantages of expressing human autoantigens in eukaryotic systems for diagnostic purposes were confirmed. |
| doi_str_mv | 10.1111/j.1365-2249.1995.tb03655.x |
| format | Article |
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Topoisomerase I (topo I) is a major systemic sclerosis (SSc)‐associated autoantigen. A cDNA construct encoding full length human topo I in a recombinant baculovirus transfer vector was used to infect insect cells in culture from which recombinant protein was purified. An ELISA using recombinant protein was evaluated in 340 sera including sera from 134 patients with SSc, of whom 33 had anti‐topo I antibodies detected by immunodiffusion. A high yield of pure topo I of expected molecular mass and catalytic activity was obtained. The recombinant topo I ELISA was 92% sensitive and 98% specific in detecting anti‐topo I antibodies which were present almost exclusively in patients with SSc. Therefore, the potential advantages of expressing human autoantigens in eukaryotic systems for diagnostic purposes were confirmed.</description><identifier>ISSN: 0009-9104</identifier><identifier>EISSN: 1365-2249</identifier><identifier>DOI: 10.1111/j.1365-2249.1995.tb03655.x</identifier><identifier>PMID: 7743657</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Animals ; Arthritis, Rheumatoid - immunology ; Autoantibodies - immunology ; DNA Topoisomerases, Type I - immunology ; Humans ; Immunoglobulin A - immunology ; Immunoglobulin M - immunology ; recombinant autoantigen ; Recombinant Proteins ; scleroderma ; Scleroderma, Systemic - immunology ; Silicosis - immunology ; Spodoptera ; topoisomerase I</subject><ispartof>Clinical and experimental immunology, 1995-05, Vol.100 (2), p.214-218</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5074-f5715fb1820b0543dc9d0aef2841ac6ed2aaba872a4db3b5beea71969b9356593</citedby><cites>FETCH-LOGICAL-c5074-f5715fb1820b0543dc9d0aef2841ac6ed2aaba872a4db3b5beea71969b9356593</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/PMC1534349/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1534349/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7743657$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>WHYTE, J.</creatorcontrib><creatorcontrib>EARNSHAW, W. C.</creatorcontrib><creatorcontrib>CHAMPOUX, J. J.</creatorcontrib><creatorcontrib>PARKER, L. H.</creatorcontrib><creatorcontrib>STEWART, L.</creatorcontrib><creatorcontrib>HALL, N. D.</creatorcontrib><creatorcontrib>MCHUGH, N. J.</creatorcontrib><title>Detection of anti‐topoisomerase I antibodies using a full length human topoisomerase I recombinant protein purified from a baculovirus expression system</title><title>Clinical and experimental immunology</title><addtitle>Clin Exp Immunol</addtitle><description>SUMMARY
Topoisomerase I (topo I) is a major systemic sclerosis (SSc)‐associated autoantigen. A cDNA construct encoding full length human topo I in a recombinant baculovirus transfer vector was used to infect insect cells in culture from which recombinant protein was purified. An ELISA using recombinant protein was evaluated in 340 sera including sera from 134 patients with SSc, of whom 33 had anti‐topo I antibodies detected by immunodiffusion. A high yield of pure topo I of expected molecular mass and catalytic activity was obtained. The recombinant topo I ELISA was 92% sensitive and 98% specific in detecting anti‐topo I antibodies which were present almost exclusively in patients with SSc. Therefore, the potential advantages of expressing human autoantigens in eukaryotic systems for diagnostic purposes were confirmed.</description><subject>Animals</subject><subject>Arthritis, Rheumatoid - immunology</subject><subject>Autoantibodies - immunology</subject><subject>DNA Topoisomerases, Type I - immunology</subject><subject>Humans</subject><subject>Immunoglobulin A - immunology</subject><subject>Immunoglobulin M - immunology</subject><subject>recombinant autoantigen</subject><subject>Recombinant Proteins</subject><subject>scleroderma</subject><subject>Scleroderma, Systemic - immunology</subject><subject>Silicosis - immunology</subject><subject>Spodoptera</subject><subject>topoisomerase I</subject><issn>0009-9104</issn><issn>1365-2249</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1995</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqVkc1u1DAUhS0EKkPhEZAsFuwSbMdOxiyQ0FDKSJW6gbVlJzczHiV2sJ0ys-MRWPfx-iRN6Gj42dUby_eec-61PoTeUJLT6bzb5bQoRcYYlzmVUuTJkKkg8v0TtDi1nqIFIURmkhL-HL2IcTc9y7JkZ-isqvgkqxbo9hMkqJP1DvsWa5fs3c9fyQ_eRt9D0BHw-nfZ-MZCxGO0boM1bseuwx24Tdri7dhrh_83Bah9b6ybzHgIPoF1eBiDbS00uA2-n1KMrsfO39gwRgz7IUCM8ybxEBP0L9GzVncRXh3vc_Tt88XX1Zfs6vpyvfp4ldWCVDxrRUVFa-iSEUMEL5paNkRDy5ac6rqEhmlt9LJimjemMMIA6IrKUhpZiFLI4hx9eMgdRtNDU4NLQXdqCLbX4aC8turfjrNbtfE3ioqCF3wOeHsMCP77CDGp3sYauk478GNUtFzyilM2Cd8_COvgYwzQnoZQomayaqdmfGrGp2ay6khW7Sfz67_XPFmPKP9844ft4PCIZLW6WDPKi3ubfbsl</recordid><startdate>199505</startdate><enddate>199505</enddate><creator>WHYTE, J.</creator><creator>EARNSHAW, W. C.</creator><creator>CHAMPOUX, J. J.</creator><creator>PARKER, L. H.</creator><creator>STEWART, L.</creator><creator>HALL, N. D.</creator><creator>MCHUGH, N. J.</creator><general>Blackwell Publishing Ltd</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>7T5</scope><scope>H94</scope><scope>5PM</scope></search><sort><creationdate>199505</creationdate><title>Detection of anti‐topoisomerase I antibodies using a full length human topoisomerase I recombinant protein purified from a baculovirus expression system</title><author>WHYTE, J. ; EARNSHAW, W. C. ; CHAMPOUX, J. J. ; PARKER, L. H. ; STEWART, L. ; HALL, N. D. ; MCHUGH, N. J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5074-f5715fb1820b0543dc9d0aef2841ac6ed2aaba872a4db3b5beea71969b9356593</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1995</creationdate><topic>Animals</topic><topic>Arthritis, Rheumatoid - immunology</topic><topic>Autoantibodies - immunology</topic><topic>DNA Topoisomerases, Type I - immunology</topic><topic>Humans</topic><topic>Immunoglobulin A - immunology</topic><topic>Immunoglobulin M - immunology</topic><topic>recombinant autoantigen</topic><topic>Recombinant Proteins</topic><topic>scleroderma</topic><topic>Scleroderma, Systemic - immunology</topic><topic>Silicosis - immunology</topic><topic>Spodoptera</topic><topic>topoisomerase I</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>WHYTE, J.</creatorcontrib><creatorcontrib>EARNSHAW, W. C.</creatorcontrib><creatorcontrib>CHAMPOUX, J. J.</creatorcontrib><creatorcontrib>PARKER, L. H.</creatorcontrib><creatorcontrib>STEWART, L.</creatorcontrib><creatorcontrib>HALL, N. D.</creatorcontrib><creatorcontrib>MCHUGH, N. J.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Immunology Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Clinical and experimental immunology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>WHYTE, J.</au><au>EARNSHAW, W. C.</au><au>CHAMPOUX, J. J.</au><au>PARKER, L. H.</au><au>STEWART, L.</au><au>HALL, N. D.</au><au>MCHUGH, N. J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection of anti‐topoisomerase I antibodies using a full length human topoisomerase I recombinant protein purified from a baculovirus expression system</atitle><jtitle>Clinical and experimental immunology</jtitle><addtitle>Clin Exp Immunol</addtitle><date>1995-05</date><risdate>1995</risdate><volume>100</volume><issue>2</issue><spage>214</spage><epage>218</epage><pages>214-218</pages><issn>0009-9104</issn><eissn>1365-2249</eissn><abstract>SUMMARY
Topoisomerase I (topo I) is a major systemic sclerosis (SSc)‐associated autoantigen. A cDNA construct encoding full length human topo I in a recombinant baculovirus transfer vector was used to infect insect cells in culture from which recombinant protein was purified. An ELISA using recombinant protein was evaluated in 340 sera including sera from 134 patients with SSc, of whom 33 had anti‐topo I antibodies detected by immunodiffusion. A high yield of pure topo I of expected molecular mass and catalytic activity was obtained. The recombinant topo I ELISA was 92% sensitive and 98% specific in detecting anti‐topo I antibodies which were present almost exclusively in patients with SSc. Therefore, the potential advantages of expressing human autoantigens in eukaryotic systems for diagnostic purposes were confirmed.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>7743657</pmid><doi>10.1111/j.1365-2249.1995.tb03655.x</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0009-9104 |
| ispartof | Clinical and experimental immunology, 1995-05, Vol.100 (2), p.214-218 |
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| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_1534349 |
| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Alma/SFX Local Collection |
| subjects | Animals Arthritis, Rheumatoid - immunology Autoantibodies - immunology DNA Topoisomerases, Type I - immunology Humans Immunoglobulin A - immunology Immunoglobulin M - immunology recombinant autoantigen Recombinant Proteins scleroderma Scleroderma, Systemic - immunology Silicosis - immunology Spodoptera topoisomerase I |
| title | Detection of anti‐topoisomerase I antibodies using a full length human topoisomerase I recombinant protein purified from a baculovirus expression system |
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