A new type of protein chip to detect hepatocellular carcinoma-related autoimmune antibodies in the sera of hepatitis C virus-positive patients
We report here a new type of protein chip to detect antibodies in sera. This chip method was used to a prototype created to detect hepatocellular carcinoma (HCC) -related autoantibodies in the sera of hepatitis C virus (HCV) infected individuals. Five cysteine-tagged (Cys-tag) and green fluorescent...
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| Veröffentlicht in: | Proteome science 2013-07, Vol.11 (1), p.33-33, Article 33 |
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| creator | Akada, Junko Kamei, Shuichi Ito, Akane Ito, Moe Kitagawa, Takao Furumoto, Hiroko Kato, Yukari Tamesa, Michiko Takashima, Motonari Shirai, Mutsunori Yamano, Hirofumi Oka, Masaaki Kuramitsu, Yasuhiro Nakamura, Kazuyuki |
| description | We report here a new type of protein chip to detect antibodies in sera. This chip method was used to a prototype created to detect hepatocellular carcinoma (HCC) -related autoantibodies in the sera of hepatitis C virus (HCV) infected individuals.
Five cysteine-tagged (Cys-tag) and green fluorescent protein (GFP)-fused recombinant heat shock protein 70 (HSP70), superoxide dismutase 2 (SOD2), and peroxiredoxin 6 (PRDX6), were spotted and immobilized on maleimide-incorporated diamond-like carbon (DLC) substrates. The antibodies in diluted sera were trapped by these proteins at each spot on the chip, and visualized by a fluorescence-conjugated anti-human IgG. The total immobilized protein level of each spot was detected with anti-GFP mouse IgG and a fluorescence-conjugated secondary anti-mouse IgG. The ratio between the two fluorescence intensities was used to quantify autoantibody levels in each serum sample. Heat treatment of the chip in a solution of denaturing and reducing agents, before serum-incubation, improved autoantibody detection. We tested serum samples from healthy individuals and HCC patients using the chips. The HSP70 autoantibodies were found at high levels in sera from HCV-positive HCC patients, but not in HCV-negative sera.
This protein chip system may have useful properties to capture a specific set of antibodies for predicting the onset of particular cancers such as HCC in HCV-infected individuals. |
| doi_str_mv | 10.1186/1477-5956-11-33 |
| format | Article |
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Five cysteine-tagged (Cys-tag) and green fluorescent protein (GFP)-fused recombinant heat shock protein 70 (HSP70), superoxide dismutase 2 (SOD2), and peroxiredoxin 6 (PRDX6), were spotted and immobilized on maleimide-incorporated diamond-like carbon (DLC) substrates. The antibodies in diluted sera were trapped by these proteins at each spot on the chip, and visualized by a fluorescence-conjugated anti-human IgG. The total immobilized protein level of each spot was detected with anti-GFP mouse IgG and a fluorescence-conjugated secondary anti-mouse IgG. The ratio between the two fluorescence intensities was used to quantify autoantibody levels in each serum sample. Heat treatment of the chip in a solution of denaturing and reducing agents, before serum-incubation, improved autoantibody detection. We tested serum samples from healthy individuals and HCC patients using the chips. The HSP70 autoantibodies were found at high levels in sera from HCV-positive HCC patients, but not in HCV-negative sera.
This protein chip system may have useful properties to capture a specific set of antibodies for predicting the onset of particular cancers such as HCC in HCV-infected individuals.</description><identifier>ISSN: 1477-5956</identifier><identifier>EISSN: 1477-5956</identifier><identifier>DOI: 10.1186/1477-5956-11-33</identifier><identifier>PMID: 23866785</identifier><language>eng</language><publisher>England: BioMed Central Ltd</publisher><subject>Analysis ; Carbon ; Cystine ; Diagnosis ; Experiments ; Health aspects ; Heat shock proteins ; Hepatitis ; Hepatitis C virus ; Hepatoma ; Immunoglobulin G ; Liver cancer ; Liver cirrhosis ; Medicine ; Methodology ; Patients ; Proteins ; Proteomics ; Risk factors ; Thiols ; Tumors ; University graduates</subject><ispartof>Proteome science, 2013-07, Vol.11 (1), p.33-33, Article 33</ispartof><rights>COPYRIGHT 2013 BioMed Central Ltd.</rights><rights>2013 Akada et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</rights><rights>Copyright © 2013 Akada et al.; licensee BioMed Central Ltd. 2013 Akada et al.; licensee BioMed Central Ltd.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-b680t-637b5e80c9a3ca62561576b3a0b7792a1572d04e8bf865e5610f8fd53bfc70053</citedby><cites>FETCH-LOGICAL-b680t-637b5e80c9a3ca62561576b3a0b7792a1572d04e8bf865e5610f8fd53bfc70053</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/PMC3750938/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3750938/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23866785$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Akada, Junko</creatorcontrib><creatorcontrib>Kamei, Shuichi</creatorcontrib><creatorcontrib>Ito, Akane</creatorcontrib><creatorcontrib>Ito, Moe</creatorcontrib><creatorcontrib>Kitagawa, Takao</creatorcontrib><creatorcontrib>Furumoto, Hiroko</creatorcontrib><creatorcontrib>Kato, Yukari</creatorcontrib><creatorcontrib>Tamesa, Michiko</creatorcontrib><creatorcontrib>Takashima, Motonari</creatorcontrib><creatorcontrib>Shirai, Mutsunori</creatorcontrib><creatorcontrib>Yamano, Hirofumi</creatorcontrib><creatorcontrib>Oka, Masaaki</creatorcontrib><creatorcontrib>Kuramitsu, Yasuhiro</creatorcontrib><creatorcontrib>Nakamura, Kazuyuki</creatorcontrib><title>A new type of protein chip to detect hepatocellular carcinoma-related autoimmune antibodies in the sera of hepatitis C virus-positive patients</title><title>Proteome science</title><addtitle>Proteome Sci</addtitle><description>We report here a new type of protein chip to detect antibodies in sera. This chip method was used to a prototype created to detect hepatocellular carcinoma (HCC) -related autoantibodies in the sera of hepatitis C virus (HCV) infected individuals.
Five cysteine-tagged (Cys-tag) and green fluorescent protein (GFP)-fused recombinant heat shock protein 70 (HSP70), superoxide dismutase 2 (SOD2), and peroxiredoxin 6 (PRDX6), were spotted and immobilized on maleimide-incorporated diamond-like carbon (DLC) substrates. The antibodies in diluted sera were trapped by these proteins at each spot on the chip, and visualized by a fluorescence-conjugated anti-human IgG. The total immobilized protein level of each spot was detected with anti-GFP mouse IgG and a fluorescence-conjugated secondary anti-mouse IgG. The ratio between the two fluorescence intensities was used to quantify autoantibody levels in each serum sample. Heat treatment of the chip in a solution of denaturing and reducing agents, before serum-incubation, improved autoantibody detection. We tested serum samples from healthy individuals and HCC patients using the chips. The HSP70 autoantibodies were found at high levels in sera from HCV-positive HCC patients, but not in HCV-negative sera.
This protein chip system may have useful properties to capture a specific set of antibodies for predicting the onset of particular cancers such as HCC in HCV-infected individuals.</description><subject>Analysis</subject><subject>Carbon</subject><subject>Cystine</subject><subject>Diagnosis</subject><subject>Experiments</subject><subject>Health aspects</subject><subject>Heat shock proteins</subject><subject>Hepatitis</subject><subject>Hepatitis C virus</subject><subject>Hepatoma</subject><subject>Immunoglobulin G</subject><subject>Liver cancer</subject><subject>Liver cirrhosis</subject><subject>Medicine</subject><subject>Methodology</subject><subject>Patients</subject><subject>Proteins</subject><subject>Proteomics</subject><subject>Risk factors</subject><subject>Thiols</subject><subject>Tumors</subject><subject>University 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hepatitis C virus-positive patients</title><author>Akada, Junko ; Kamei, Shuichi ; Ito, Akane ; Ito, Moe ; Kitagawa, Takao ; Furumoto, Hiroko ; Kato, Yukari ; Tamesa, Michiko ; Takashima, Motonari ; Shirai, Mutsunori ; Yamano, Hirofumi ; Oka, Masaaki ; Kuramitsu, Yasuhiro ; Nakamura, Kazuyuki</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-b680t-637b5e80c9a3ca62561576b3a0b7792a1572d04e8bf865e5610f8fd53bfc70053</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Analysis</topic><topic>Carbon</topic><topic>Cystine</topic><topic>Diagnosis</topic><topic>Experiments</topic><topic>Health aspects</topic><topic>Heat shock proteins</topic><topic>Hepatitis</topic><topic>Hepatitis C virus</topic><topic>Hepatoma</topic><topic>Immunoglobulin G</topic><topic>Liver cancer</topic><topic>Liver 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autoimmune antibodies in the sera of hepatitis C virus-positive patients</atitle><jtitle>Proteome science</jtitle><addtitle>Proteome Sci</addtitle><date>2013-07-19</date><risdate>2013</risdate><volume>11</volume><issue>1</issue><spage>33</spage><epage>33</epage><pages>33-33</pages><artnum>33</artnum><issn>1477-5956</issn><eissn>1477-5956</eissn><abstract>We report here a new type of protein chip to detect antibodies in sera. This chip method was used to a prototype created to detect hepatocellular carcinoma (HCC) -related autoantibodies in the sera of hepatitis C virus (HCV) infected individuals.
Five cysteine-tagged (Cys-tag) and green fluorescent protein (GFP)-fused recombinant heat shock protein 70 (HSP70), superoxide dismutase 2 (SOD2), and peroxiredoxin 6 (PRDX6), were spotted and immobilized on maleimide-incorporated diamond-like carbon (DLC) substrates. The antibodies in diluted sera were trapped by these proteins at each spot on the chip, and visualized by a fluorescence-conjugated anti-human IgG. The total immobilized protein level of each spot was detected with anti-GFP mouse IgG and a fluorescence-conjugated secondary anti-mouse IgG. The ratio between the two fluorescence intensities was used to quantify autoantibody levels in each serum sample. Heat treatment of the chip in a solution of denaturing and reducing agents, before serum-incubation, improved autoantibody detection. We tested serum samples from healthy individuals and HCC patients using the chips. The HSP70 autoantibodies were found at high levels in sera from HCV-positive HCC patients, but not in HCV-negative sera.
This protein chip system may have useful properties to capture a specific set of antibodies for predicting the onset of particular cancers such as HCC in HCV-infected individuals.</abstract><cop>England</cop><pub>BioMed Central Ltd</pub><pmid>23866785</pmid><doi>10.1186/1477-5956-11-33</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Analysis Carbon Cystine Diagnosis Experiments Health aspects Heat shock proteins Hepatitis Hepatitis C virus Hepatoma Immunoglobulin G Liver cancer Liver cirrhosis Medicine Methodology Patients Proteins Proteomics Risk factors Thiols Tumors University graduates |
| title | A new type of protein chip to detect hepatocellular carcinoma-related autoimmune antibodies in the sera of hepatitis C virus-positive patients |
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