An enzyme-linked immunosorbent assay for the detection of hepatocyte plasma membrane antibodies
An enzyme-linked immunosorbent assay (ELISA) using plates coated with hepatocyte plasma membranes (HPM) was developed for the measurement of antibodies directed at hepatocyte surface antigens. Precoating ELISA plates with poly- L-lysine (PLL) provided firm attachement for the adsorption of HPM. The...
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| Veröffentlicht in: | Journal of immunological methods 1985-12, Vol.85 (1), p.203-216 |
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| creator | Swanson, N.R. Bartholomeus, W.N. Reed, W.D. Joske, R.A. |
| description | An enzyme-linked immunosorbent assay (ELISA) using plates coated with hepatocyte plasma membranes (HPM) was developed for the measurement of antibodies directed at hepatocyte surface antigens. Precoating ELISA plates with poly-
L-lysine (PLL) provided firm attachement for the adsorption of HPM. The use of HPM, in preference to whole hepatocytes, excludes pathologically irrelevant cytoplasmic antigens. In addition, there is no necessity for glutaraldehyde fixation which is commonly used in cellular assays to maintain cellular integrity and which may result in loss or alteration in antigenic specificities.
The assay was used to study loss of tolerance to mouse HPM in mice immunized with rat HPM. Three mouse strains were immunized, each strain developed antibodies to rat HPM and autoantibodies to mouse HPM with autoantibody levels reaching a peak 6–10 weeks after commencement of immunization. The correlation between ELISA and indirect immunofluorescence for the measurement of HPM autoantibodies was 0.79 (P < 0.001) within the serum titration range of 1:25 to 1:200
Antibody to control kidney plasma membrane (KPM) was also measured by ELISA, after elimination of endogenous alkaline phosphatase activity using levamisole. Immunization with rat HPM elicited organ-non-specific autoantibodies to KPM, but these were at lower levels than autoantibodies to HPM. |
| doi_str_mv | 10.1016/0022-1759(85)90288-1 |
| format | Article |
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L-lysine (PLL) provided firm attachement for the adsorption of HPM. The use of HPM, in preference to whole hepatocytes, excludes pathologically irrelevant cytoplasmic antigens. In addition, there is no necessity for glutaraldehyde fixation which is commonly used in cellular assays to maintain cellular integrity and which may result in loss or alteration in antigenic specificities.
The assay was used to study loss of tolerance to mouse HPM in mice immunized with rat HPM. Three mouse strains were immunized, each strain developed antibodies to rat HPM and autoantibodies to mouse HPM with autoantibody levels reaching a peak 6–10 weeks after commencement of immunization. The correlation between ELISA and indirect immunofluorescence for the measurement of HPM autoantibodies was 0.79 (P < 0.001) within the serum titration range of 1:25 to 1:200
Antibody to control kidney plasma membrane (KPM) was also measured by ELISA, after elimination of endogenous alkaline phosphatase activity using levamisole. Immunization with rat HPM elicited organ-non-specific autoantibodies to KPM, but these were at lower levels than autoantibodies to HPM.</description><identifier>ISSN: 0022-1759</identifier><identifier>EISSN: 1872-7905</identifier><identifier>DOI: 10.1016/0022-1759(85)90288-1</identifier><identifier>PMID: 3908561</identifier><identifier>CODEN: JIMMBG</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Alkaline Phosphatase - metabolism ; Animals ; autoantibodies ; Autoantibodies - analysis ; Biological and medical sciences ; Cell Membrane - immunology ; Dose-Response Relationship, Immunologic ; ELISA ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technique ; Fundamental and applied biological sciences. Psychology ; Fundamental immunology ; hepatocyte plasma membrane ; immunofluoresce ; Liver - immunology ; Mice ; Mice, Inbred Strains ; Molecular immunology ; mouse ; Polylysine ; Rats ; Techniques</subject><ispartof>Journal of immunological methods, 1985-12, Vol.85 (1), p.203-216</ispartof><rights>1985</rights><rights>1986 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c386t-80589df9cd67cc8a835e0af73ed759b13eecabbca10e2d03e6120be611b8bb9f3</citedby><cites>FETCH-LOGICAL-c386t-80589df9cd67cc8a835e0af73ed759b13eecabbca10e2d03e6120be611b8bb9f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0022-1759(85)90288-1$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=8751838$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/3908561$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Swanson, N.R.</creatorcontrib><creatorcontrib>Bartholomeus, W.N.</creatorcontrib><creatorcontrib>Reed, W.D.</creatorcontrib><creatorcontrib>Joske, R.A.</creatorcontrib><title>An enzyme-linked immunosorbent assay for the detection of hepatocyte plasma membrane antibodies</title><title>Journal of immunological methods</title><addtitle>J Immunol Methods</addtitle><description>An enzyme-linked immunosorbent assay (ELISA) using plates coated with hepatocyte plasma membranes (HPM) was developed for the measurement of antibodies directed at hepatocyte surface antigens. Precoating ELISA plates with poly-
L-lysine (PLL) provided firm attachement for the adsorption of HPM. The use of HPM, in preference to whole hepatocytes, excludes pathologically irrelevant cytoplasmic antigens. In addition, there is no necessity for glutaraldehyde fixation which is commonly used in cellular assays to maintain cellular integrity and which may result in loss or alteration in antigenic specificities.
The assay was used to study loss of tolerance to mouse HPM in mice immunized with rat HPM. Three mouse strains were immunized, each strain developed antibodies to rat HPM and autoantibodies to mouse HPM with autoantibody levels reaching a peak 6–10 weeks after commencement of immunization. The correlation between ELISA and indirect immunofluorescence for the measurement of HPM autoantibodies was 0.79 (P < 0.001) within the serum titration range of 1:25 to 1:200
Antibody to control kidney plasma membrane (KPM) was also measured by ELISA, after elimination of endogenous alkaline phosphatase activity using levamisole. Immunization with rat HPM elicited organ-non-specific autoantibodies to KPM, but these were at lower levels than autoantibodies to HPM.</description><subject>Alkaline Phosphatase - metabolism</subject><subject>Animals</subject><subject>autoantibodies</subject><subject>Autoantibodies - analysis</subject><subject>Biological and medical sciences</subject><subject>Cell Membrane - immunology</subject><subject>Dose-Response Relationship, Immunologic</subject><subject>ELISA</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Fluorescent Antibody Technique</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fundamental immunology</subject><subject>hepatocyte plasma membrane</subject><subject>immunofluoresce</subject><subject>Liver - immunology</subject><subject>Mice</subject><subject>Mice, Inbred Strains</subject><subject>Molecular immunology</subject><subject>mouse</subject><subject>Polylysine</subject><subject>Rats</subject><subject>Techniques</subject><issn>0022-1759</issn><issn>1872-7905</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1985</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE2LFDEQhoMo6-zqP1DIQWQ9tFY6pju5LCyLX7DgRc8hH9VstJOMSUYYf70ZZ5ijl6pDPW_x8hDygsFbBmx6BzCOA5uFupbijYJRyoE9Ihsm53GYFYjHZHNGnpLLWn8AAIMJLsgFVyDFxDZE3yaK6c8-4rCG9BM9DTHuUq65WEyNmlrNni650PaA1GND10JONC_0AbemZbdvSLerqdHQiNEWk5Ca1ILNPmB9Rp4sZq34_LSvyPePH77dfR7uv376cnd7PzgupzZIEFL5RTk_zc5JI7lAMMvM0ff2lnFEZ6x1hgGOHjhObATbJ7PSWrXwK_L6-Hdb8q8d1qZjqA7XtdfJu6rnSXAlmerg-yPoSq614KK3JURT9pqBPnjVB2n6IE1Lof951azHXp7-72xEfw6dRPb7q9PdVGfWpWtwoZ4xOQsmuezYzRHD7uJ3wKKrC5gc-lC6We1z-H-Pv1y9lhM</recordid><startdate>19851217</startdate><enddate>19851217</enddate><creator>Swanson, N.R.</creator><creator>Bartholomeus, W.N.</creator><creator>Reed, W.D.</creator><creator>Joske, R.A.</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><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>7X8</scope></search><sort><creationdate>19851217</creationdate><title>An enzyme-linked immunosorbent assay for the detection of hepatocyte plasma membrane antibodies</title><author>Swanson, N.R. ; Bartholomeus, W.N. ; Reed, W.D. ; Joske, R.A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c386t-80589df9cd67cc8a835e0af73ed759b13eecabbca10e2d03e6120be611b8bb9f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1985</creationdate><topic>Alkaline Phosphatase - metabolism</topic><topic>Animals</topic><topic>autoantibodies</topic><topic>Autoantibodies - analysis</topic><topic>Biological and medical sciences</topic><topic>Cell Membrane - immunology</topic><topic>Dose-Response Relationship, Immunologic</topic><topic>ELISA</topic><topic>Enzyme-Linked Immunosorbent Assay</topic><topic>Fluorescent Antibody Technique</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fundamental immunology</topic><topic>hepatocyte plasma membrane</topic><topic>immunofluoresce</topic><topic>Liver - immunology</topic><topic>Mice</topic><topic>Mice, Inbred Strains</topic><topic>Molecular immunology</topic><topic>mouse</topic><topic>Polylysine</topic><topic>Rats</topic><topic>Techniques</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Swanson, N.R.</creatorcontrib><creatorcontrib>Bartholomeus, W.N.</creatorcontrib><creatorcontrib>Reed, W.D.</creatorcontrib><creatorcontrib>Joske, R.A.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of immunological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Swanson, N.R.</au><au>Bartholomeus, W.N.</au><au>Reed, W.D.</au><au>Joske, R.A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>An enzyme-linked immunosorbent assay for the detection of hepatocyte plasma membrane antibodies</atitle><jtitle>Journal of immunological methods</jtitle><addtitle>J Immunol Methods</addtitle><date>1985-12-17</date><risdate>1985</risdate><volume>85</volume><issue>1</issue><spage>203</spage><epage>216</epage><pages>203-216</pages><issn>0022-1759</issn><eissn>1872-7905</eissn><coden>JIMMBG</coden><abstract>An enzyme-linked immunosorbent assay (ELISA) using plates coated with hepatocyte plasma membranes (HPM) was developed for the measurement of antibodies directed at hepatocyte surface antigens. Precoating ELISA plates with poly-
L-lysine (PLL) provided firm attachement for the adsorption of HPM. The use of HPM, in preference to whole hepatocytes, excludes pathologically irrelevant cytoplasmic antigens. In addition, there is no necessity for glutaraldehyde fixation which is commonly used in cellular assays to maintain cellular integrity and which may result in loss or alteration in antigenic specificities.
The assay was used to study loss of tolerance to mouse HPM in mice immunized with rat HPM. Three mouse strains were immunized, each strain developed antibodies to rat HPM and autoantibodies to mouse HPM with autoantibody levels reaching a peak 6–10 weeks after commencement of immunization. The correlation between ELISA and indirect immunofluorescence for the measurement of HPM autoantibodies was 0.79 (P < 0.001) within the serum titration range of 1:25 to 1:200
Antibody to control kidney plasma membrane (KPM) was also measured by ELISA, after elimination of endogenous alkaline phosphatase activity using levamisole. Immunization with rat HPM elicited organ-non-specific autoantibodies to KPM, but these were at lower levels than autoantibodies to HPM.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>3908561</pmid><doi>10.1016/0022-1759(85)90288-1</doi><tpages>14</tpages></addata></record> |
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| subjects | Alkaline Phosphatase - metabolism Animals autoantibodies Autoantibodies - analysis Biological and medical sciences Cell Membrane - immunology Dose-Response Relationship, Immunologic ELISA Enzyme-Linked Immunosorbent Assay Fluorescent Antibody Technique Fundamental and applied biological sciences. Psychology Fundamental immunology hepatocyte plasma membrane immunofluoresce Liver - immunology Mice Mice, Inbred Strains Molecular immunology mouse Polylysine Rats Techniques |
| title | An enzyme-linked immunosorbent assay for the detection of hepatocyte plasma membrane antibodies |
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