Platelet-Collagen Adhesion: Inhibition by a Monoclonal Antibody That Binds Glycoprotein IIb
To identify platelet surface structures involved in adhesion to collagen, the effect of 16 murine antiplatelet membrane hybridoma antibodies were tested in a defined, in vitro assay. Four of these antibodies inhibited platelet-collagen adhesion and reacted with a polypeptide with Mr ∼ 125,000, as de...
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| Veröffentlicht in: | The Journal of cell biology 1984-12, Vol.99 (6), p.2056-2060 |
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| creator | Shadle, Paula J. Ginsberg, Mark H. Plow, Edward F. Barondes, Samuel H. |
| description | To identify platelet surface structures involved in adhesion to collagen, the effect of 16 murine antiplatelet membrane hybridoma antibodies were tested in a defined, in vitro assay. Four of these antibodies inhibited platelet-collagen adhesion and reacted with a polypeptide with Mr ∼ 125,000, as determined by immunoblots after gel electrophoresis under reducing conditions. Through detailed studies with one of these antibodies, the monoclonal antibody PMI-1, the relevant antigen was identified as platelet glycoprotein IIbα, based upon (a) co-migration with this glycoprotein in two-dimensional gel electrophoresis and (b) co-purification by immunoaffinity chromatography with a protein with apparent Mr identical to that of glycoprotein III, under conditions in which glycoproteins IIb and III form a complex. Univalent antibody fragments prepared from monoclonal antibody PMI-1 inhibited >80% of platelet-collagen adhesion, and inhibition was completely blocked by the immunopurified antigen. These results indicate that glycoprotein IIb participates in some aspect of platelet-collagen adhesion. In contrast, the purified antigen only partially neutralized a polyclonal antiserum that blocked platelet-collagen adhesion, to a maximum of ∼25%, at saturating antigen concentrations. Thus, by these immunological criteria, glycoprotein IIb is not the only molecule involved in this process. |
| doi_str_mv | 10.1083/jcb.99.6.2056 |
| format | Article |
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Four of these antibodies inhibited platelet-collagen adhesion and reacted with a polypeptide with Mr ∼ 125,000, as determined by immunoblots after gel electrophoresis under reducing conditions. Through detailed studies with one of these antibodies, the monoclonal antibody PMI-1, the relevant antigen was identified as platelet glycoprotein IIbα, based upon (a) co-migration with this glycoprotein in two-dimensional gel electrophoresis and (b) co-purification by immunoaffinity chromatography with a protein with apparent Mr identical to that of glycoprotein III, under conditions in which glycoproteins IIb and III form a complex. Univalent antibody fragments prepared from monoclonal antibody PMI-1 inhibited >80% of platelet-collagen adhesion, and inhibition was completely blocked by the immunopurified antigen. These results indicate that glycoprotein IIb participates in some aspect of platelet-collagen adhesion. In contrast, the purified antigen only partially neutralized a polyclonal antiserum that blocked platelet-collagen adhesion, to a maximum of ∼25%, at saturating antigen concentrations. Thus, by these immunological criteria, glycoprotein IIb is not the only molecule involved in this process.</description><identifier>ISSN: 0021-9525</identifier><identifier>EISSN: 1540-8140</identifier><identifier>DOI: 10.1083/jcb.99.6.2056</identifier><identifier>PMID: 6238974</identifier><identifier>CODEN: JCLBA3</identifier><language>eng</language><publisher>New York, NY: Rockefeller University Press</publisher><subject>Adhesion ; Antibodies ; Antibodies, Monoclonal ; Antigen-Antibody Complex ; Antigens ; Antiserum ; Biological and medical sciences ; Blood coagulation. Blood cells ; Blood Platelets - physiology ; Chromatography, Affinity ; Collagen ; Collagens ; Electrophoresis, Polyacrylamide Gel ; Fundamental and applied biological sciences. Psychology ; Gels ; Glycoproteins ; Glycoproteins - analysis ; Humans ; Hybridomas ; Membrane Proteins - analysis ; Molecular and cellular biology ; Molecular Weight ; Monoclonal antibodies ; Platelet ; Platelet Aggregation ; Platelet Membrane Glycoproteins ; Platelets ; Radioimmunoassay</subject><ispartof>The Journal of cell biology, 1984-12, Vol.99 (6), p.2056-2060</ispartof><rights>Copyright 1984 The Rockefeller University Press</rights><rights>1985 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c432t-f2ce671fbe03588e8f699f3439136fab50fba9a9946ee9fd0c3fb4d0e965f2f53</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=9124255$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/6238974$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Shadle, Paula J.</creatorcontrib><creatorcontrib>Ginsberg, Mark H.</creatorcontrib><creatorcontrib>Plow, Edward F.</creatorcontrib><creatorcontrib>Barondes, Samuel H.</creatorcontrib><title>Platelet-Collagen Adhesion: Inhibition by a Monoclonal Antibody That Binds Glycoprotein IIb</title><title>The Journal of cell biology</title><addtitle>J Cell Biol</addtitle><description>To identify platelet surface structures involved in adhesion to collagen, the effect of 16 murine antiplatelet membrane hybridoma antibodies were tested in a defined, in vitro assay. Four of these antibodies inhibited platelet-collagen adhesion and reacted with a polypeptide with Mr ∼ 125,000, as determined by immunoblots after gel electrophoresis under reducing conditions. Through detailed studies with one of these antibodies, the monoclonal antibody PMI-1, the relevant antigen was identified as platelet glycoprotein IIbα, based upon (a) co-migration with this glycoprotein in two-dimensional gel electrophoresis and (b) co-purification by immunoaffinity chromatography with a protein with apparent Mr identical to that of glycoprotein III, under conditions in which glycoproteins IIb and III form a complex. Univalent antibody fragments prepared from monoclonal antibody PMI-1 inhibited >80% of platelet-collagen adhesion, and inhibition was completely blocked by the immunopurified antigen. These results indicate that glycoprotein IIb participates in some aspect of platelet-collagen adhesion. In contrast, the purified antigen only partially neutralized a polyclonal antiserum that blocked platelet-collagen adhesion, to a maximum of ∼25%, at saturating antigen concentrations. Thus, by these immunological criteria, glycoprotein IIb is not the only molecule involved in this process.</description><subject>Adhesion</subject><subject>Antibodies</subject><subject>Antibodies, Monoclonal</subject><subject>Antigen-Antibody Complex</subject><subject>Antigens</subject><subject>Antiserum</subject><subject>Biological and medical sciences</subject><subject>Blood coagulation. Blood cells</subject><subject>Blood Platelets - physiology</subject><subject>Chromatography, Affinity</subject><subject>Collagen</subject><subject>Collagens</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gels</subject><subject>Glycoproteins</subject><subject>Glycoproteins - analysis</subject><subject>Humans</subject><subject>Hybridomas</subject><subject>Membrane Proteins - analysis</subject><subject>Molecular and cellular biology</subject><subject>Molecular Weight</subject><subject>Monoclonal antibodies</subject><subject>Platelet</subject><subject>Platelet Aggregation</subject><subject>Platelet Membrane Glycoproteins</subject><subject>Platelets</subject><subject>Radioimmunoassay</subject><issn>0021-9525</issn><issn>1540-8140</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1984</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkc2LUzEUxYMoY2d06U4hC3H3ar7fiwuhlnEsjOhiXLkISV4yTUmTmqRC_3tfaam6uhfOj3Mu9wDwCqM5RgN9v7FmLuVczAni4gmYYc5QN2CGnoIZQgR3khP-HFzXukEIsZ7RK3AlCB1kz2bg5_eom4uudcsco350CS7Gtashpw9wldbBhDbt0Byghl9zyjbmpCNcpBZMHg_wYa0b_BTSWOFdPNi8K7m5kOBqZV6AZ17H6l6e5w348fn2Yfmlu_92t1ou7jvLKGmdJ9aJHnvjEOXD4AYvpPSUUYmp8Npw5I2WWkomnJN-RJZ6w0bkpOCeeE5vwMeT725vtm60LrWio9qVsNXloLIO6n8lhbV6zL8VwZhygSeDd2eDkn_tXW1qG6p10z-Sy_uqet6LAaNjUncCbcm1FucvIRipYxtqakNJqYQ6tjHxb_697EKf3z_pb8-6rlZHX3SyoV4wiQkj_Bj7-oRtasvlb6bASPaI_gG2uZ2k</recordid><startdate>19841201</startdate><enddate>19841201</enddate><creator>Shadle, Paula J.</creator><creator>Ginsberg, Mark H.</creator><creator>Plow, Edward F.</creator><creator>Barondes, Samuel H.</creator><general>Rockefeller University Press</general><general>The Rockefeller University Press</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><scope>5PM</scope></search><sort><creationdate>19841201</creationdate><title>Platelet-Collagen Adhesion: Inhibition by a Monoclonal Antibody That Binds Glycoprotein IIb</title><author>Shadle, Paula J. ; Ginsberg, Mark H. ; Plow, Edward F. ; Barondes, Samuel H.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c432t-f2ce671fbe03588e8f699f3439136fab50fba9a9946ee9fd0c3fb4d0e965f2f53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1984</creationdate><topic>Adhesion</topic><topic>Antibodies</topic><topic>Antibodies, Monoclonal</topic><topic>Antigen-Antibody Complex</topic><topic>Antigens</topic><topic>Antiserum</topic><topic>Biological and medical sciences</topic><topic>Blood coagulation. Blood cells</topic><topic>Blood Platelets - physiology</topic><topic>Chromatography, Affinity</topic><topic>Collagen</topic><topic>Collagens</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gels</topic><topic>Glycoproteins</topic><topic>Glycoproteins - analysis</topic><topic>Humans</topic><topic>Hybridomas</topic><topic>Membrane Proteins - analysis</topic><topic>Molecular and cellular biology</topic><topic>Molecular Weight</topic><topic>Monoclonal antibodies</topic><topic>Platelet</topic><topic>Platelet Aggregation</topic><topic>Platelet Membrane Glycoproteins</topic><topic>Platelets</topic><topic>Radioimmunoassay</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Shadle, Paula J.</creatorcontrib><creatorcontrib>Ginsberg, Mark H.</creatorcontrib><creatorcontrib>Plow, Edward F.</creatorcontrib><creatorcontrib>Barondes, Samuel H.</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><collection>PubMed Central (Full Participant titles)</collection><jtitle>The Journal of cell biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Shadle, Paula J.</au><au>Ginsberg, Mark H.</au><au>Plow, Edward F.</au><au>Barondes, Samuel H.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Platelet-Collagen Adhesion: Inhibition by a Monoclonal Antibody That Binds Glycoprotein IIb</atitle><jtitle>The Journal of cell biology</jtitle><addtitle>J Cell Biol</addtitle><date>1984-12-01</date><risdate>1984</risdate><volume>99</volume><issue>6</issue><spage>2056</spage><epage>2060</epage><pages>2056-2060</pages><issn>0021-9525</issn><eissn>1540-8140</eissn><coden>JCLBA3</coden><abstract>To identify platelet surface structures involved in adhesion to collagen, the effect of 16 murine antiplatelet membrane hybridoma antibodies were tested in a defined, in vitro assay. Four of these antibodies inhibited platelet-collagen adhesion and reacted with a polypeptide with Mr ∼ 125,000, as determined by immunoblots after gel electrophoresis under reducing conditions. Through detailed studies with one of these antibodies, the monoclonal antibody PMI-1, the relevant antigen was identified as platelet glycoprotein IIbα, based upon (a) co-migration with this glycoprotein in two-dimensional gel electrophoresis and (b) co-purification by immunoaffinity chromatography with a protein with apparent Mr identical to that of glycoprotein III, under conditions in which glycoproteins IIb and III form a complex. Univalent antibody fragments prepared from monoclonal antibody PMI-1 inhibited >80% of platelet-collagen adhesion, and inhibition was completely blocked by the immunopurified antigen. These results indicate that glycoprotein IIb participates in some aspect of platelet-collagen adhesion. In contrast, the purified antigen only partially neutralized a polyclonal antiserum that blocked platelet-collagen adhesion, to a maximum of ∼25%, at saturating antigen concentrations. Thus, by these immunological criteria, glycoprotein IIb is not the only molecule involved in this process.</abstract><cop>New York, NY</cop><pub>Rockefeller University Press</pub><pmid>6238974</pmid><doi>10.1083/jcb.99.6.2056</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Adhesion Antibodies Antibodies, Monoclonal Antigen-Antibody Complex Antigens Antiserum Biological and medical sciences Blood coagulation. Blood cells Blood Platelets - physiology Chromatography, Affinity Collagen Collagens Electrophoresis, Polyacrylamide Gel Fundamental and applied biological sciences. Psychology Gels Glycoproteins Glycoproteins - analysis Humans Hybridomas Membrane Proteins - analysis Molecular and cellular biology Molecular Weight Monoclonal antibodies Platelet Platelet Aggregation Platelet Membrane Glycoproteins Platelets Radioimmunoassay |
| title | Platelet-Collagen Adhesion: Inhibition by a Monoclonal Antibody That Binds Glycoprotein IIb |
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