Generation of murine monoclonal antihuman milk fat globule membrane antibodies using immunoprecipitation and BIAcore analyses
A selection of monoclonal antibodies was developed against deoxycholine-solubilized human milk fat globule membranes (HMFG). The antibodies were selected for their ability to immunoprecipitate 125I-labeled HMFG and then further analyzed by surface plasmon resonance on the BIAcore for their reactivit...
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| Veröffentlicht in: | Hybridoma 1995-12, Vol.14 (6), p.587-591 |
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| creator | BYNUM, J HUTCHINS, J. T KULL, F. C |
| description | A selection of monoclonal antibodies was developed against deoxycholine-solubilized human milk fat globule membranes (HMFG). The antibodies were selected for their ability to immunoprecipitate 125I-labeled HMFG and then further analyzed by surface plasmon resonance on the BIAcore for their reactivity with HMFG and with a fusion protein containing a 4-mer of the muc-1 tandem repeat. Both the HMFG and the fusion protein proved to be robust surfaces for the analysis of crude supernatants. The BIAcore evaluation was useful in identifying true positives. BIAcore analyses of purified antibody preparations were used to determine binding characteristics such as affinity and intensity. The latter proved useful in selecting a panel for evaluation by immunohistochemistry for breast tissue reactivity. Four of 6 antibodies appeared to react more intensely with tumor compared with normal breast tissues. One of those antibodies reacted with the fusion protein 4-mer of the muc-1 tandem repeat. |
| doi_str_mv | 10.1089/hyb.1995.14.587 |
| format | Article |
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T ; KULL, F. C</creator><creatorcontrib>BYNUM, J ; HUTCHINS, J. T ; KULL, F. C</creatorcontrib><description>A selection of monoclonal antibodies was developed against deoxycholine-solubilized human milk fat globule membranes (HMFG). The antibodies were selected for their ability to immunoprecipitate 125I-labeled HMFG and then further analyzed by surface plasmon resonance on the BIAcore for their reactivity with HMFG and with a fusion protein containing a 4-mer of the muc-1 tandem repeat. Both the HMFG and the fusion protein proved to be robust surfaces for the analysis of crude supernatants. The BIAcore evaluation was useful in identifying true positives. BIAcore analyses of purified antibody preparations were used to determine binding characteristics such as affinity and intensity. The latter proved useful in selecting a panel for evaluation by immunohistochemistry for breast tissue reactivity. Four of 6 antibodies appeared to react more intensely with tumor compared with normal breast tissues. One of those antibodies reacted with the fusion protein 4-mer of the muc-1 tandem repeat.</description><identifier>ISSN: 0272-457X</identifier><identifier>EISSN: 2168-7897</identifier><identifier>EISSN: 0272-457X</identifier><identifier>DOI: 10.1089/hyb.1995.14.587</identifier><identifier>PMID: 8770647</identifier><identifier>CODEN: HYBRDY</identifier><language>eng</language><publisher>Larchmont, NY: Liebert</publisher><subject>Animals ; Antibodies, immunoglobulins ; Antibodies, Monoclonal - biosynthesis ; Antibody Affinity ; Biological and medical sciences ; Biosensing Techniques ; Female ; Fundamental and applied biological sciences. Psychology ; Fundamental immunology ; Humans ; Hybridomas - chemistry ; Hybridomas - metabolism ; Membrane Glycoproteins - immunology ; Mice ; Mice, Inbred BALB C ; Milk, Human - immunology ; Molecular immunology ; Monoclonal antibodies ; Mucin-1 - immunology ; Precipitin Tests</subject><ispartof>Hybridoma, 1995-12, Vol.14 (6), p.587-591</ispartof><rights>1996 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c275t-43259a4a18652c637955956d7111de44c4a9e967af401f73082d286f19de10773</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,3028,27903,27904</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=2974978$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8770647$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>BYNUM, J</creatorcontrib><creatorcontrib>HUTCHINS, J. T</creatorcontrib><creatorcontrib>KULL, F. C</creatorcontrib><title>Generation of murine monoclonal antihuman milk fat globule membrane antibodies using immunoprecipitation and BIAcore analyses</title><title>Hybridoma</title><addtitle>Hybridoma</addtitle><description>A selection of monoclonal antibodies was developed against deoxycholine-solubilized human milk fat globule membranes (HMFG). The antibodies were selected for their ability to immunoprecipitate 125I-labeled HMFG and then further analyzed by surface plasmon resonance on the BIAcore for their reactivity with HMFG and with a fusion protein containing a 4-mer of the muc-1 tandem repeat. Both the HMFG and the fusion protein proved to be robust surfaces for the analysis of crude supernatants. The BIAcore evaluation was useful in identifying true positives. BIAcore analyses of purified antibody preparations were used to determine binding characteristics such as affinity and intensity. The latter proved useful in selecting a panel for evaluation by immunohistochemistry for breast tissue reactivity. Four of 6 antibodies appeared to react more intensely with tumor compared with normal breast tissues. One of those antibodies reacted with the fusion protein 4-mer of the muc-1 tandem repeat.</description><subject>Animals</subject><subject>Antibodies, immunoglobulins</subject><subject>Antibodies, Monoclonal - biosynthesis</subject><subject>Antibody Affinity</subject><subject>Biological and medical sciences</subject><subject>Biosensing Techniques</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fundamental immunology</subject><subject>Humans</subject><subject>Hybridomas - chemistry</subject><subject>Hybridomas - metabolism</subject><subject>Membrane Glycoproteins - immunology</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>Milk, Human - immunology</subject><subject>Molecular immunology</subject><subject>Monoclonal antibodies</subject><subject>Mucin-1 - immunology</subject><subject>Precipitin Tests</subject><issn>0272-457X</issn><issn>2168-7897</issn><issn>0272-457X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1995</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9kDtvHCEUhZGVyFk7qV1FoojSzRoYmAulY8UPyZKbREqHGIaxSXhsYKbYIv89rHbl6hbnO0dXH0JXlGwpker6dT9uqVJiS_lWSDhDG0YH2YFU8A5tCAPWcQG_PqCLWn8TQgTjcI7OJQAZOGzQv3uXXDGLzwnnGce1-ORwzCnbkJMJ2KTFv67RJBx9-INns-CXkMc1NMrFsZiGH5gxT95VvFafXrCPcU15V5z1O78c102a8LfHG5vLoWDCvrr6Eb2fTaju0-leop9333_cPnRPz_ePtzdPnWUglo73TCjDDZWDYHboQQmhxDABpXRynFtulFMDmJkTOkNPJJuYHGaqJkcJQH-Jvh53dyX_XV1ddPTVuhDa93mtGpovArRv4PURtCXXWtysd8VHU_aaEn0QrptwfRCuKddNeGt8Pk2vY3TTG38y3PIvp9xUa8LchFlf3zCmgCuQ_X8J_Irm</recordid><startdate>19951201</startdate><enddate>19951201</enddate><creator>BYNUM, J</creator><creator>HUTCHINS, J. T</creator><creator>KULL, F. C</creator><general>Liebert</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>19951201</creationdate><title>Generation of murine monoclonal antihuman milk fat globule membrane antibodies using immunoprecipitation and BIAcore analyses</title><author>BYNUM, J ; HUTCHINS, J. T ; KULL, F. C</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c275t-43259a4a18652c637955956d7111de44c4a9e967af401f73082d286f19de10773</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1995</creationdate><topic>Animals</topic><topic>Antibodies, immunoglobulins</topic><topic>Antibodies, Monoclonal - biosynthesis</topic><topic>Antibody Affinity</topic><topic>Biological and medical sciences</topic><topic>Biosensing Techniques</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fundamental immunology</topic><topic>Humans</topic><topic>Hybridomas - chemistry</topic><topic>Hybridomas - metabolism</topic><topic>Membrane Glycoproteins - immunology</topic><topic>Mice</topic><topic>Mice, Inbred BALB C</topic><topic>Milk, Human - immunology</topic><topic>Molecular immunology</topic><topic>Monoclonal antibodies</topic><topic>Mucin-1 - immunology</topic><topic>Precipitin Tests</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>BYNUM, J</creatorcontrib><creatorcontrib>HUTCHINS, J. T</creatorcontrib><creatorcontrib>KULL, F. C</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>Hybridoma</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>BYNUM, J</au><au>HUTCHINS, J. T</au><au>KULL, F. C</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Generation of murine monoclonal antihuman milk fat globule membrane antibodies using immunoprecipitation and BIAcore analyses</atitle><jtitle>Hybridoma</jtitle><addtitle>Hybridoma</addtitle><date>1995-12-01</date><risdate>1995</risdate><volume>14</volume><issue>6</issue><spage>587</spage><epage>591</epage><pages>587-591</pages><issn>0272-457X</issn><eissn>2168-7897</eissn><eissn>0272-457X</eissn><coden>HYBRDY</coden><abstract>A selection of monoclonal antibodies was developed against deoxycholine-solubilized human milk fat globule membranes (HMFG). The antibodies were selected for their ability to immunoprecipitate 125I-labeled HMFG and then further analyzed by surface plasmon resonance on the BIAcore for their reactivity with HMFG and with a fusion protein containing a 4-mer of the muc-1 tandem repeat. Both the HMFG and the fusion protein proved to be robust surfaces for the analysis of crude supernatants. The BIAcore evaluation was useful in identifying true positives. BIAcore analyses of purified antibody preparations were used to determine binding characteristics such as affinity and intensity. The latter proved useful in selecting a panel for evaluation by immunohistochemistry for breast tissue reactivity. Four of 6 antibodies appeared to react more intensely with tumor compared with normal breast tissues. One of those antibodies reacted with the fusion protein 4-mer of the muc-1 tandem repeat.</abstract><cop>Larchmont, NY</cop><pub>Liebert</pub><pmid>8770647</pmid><doi>10.1089/hyb.1995.14.587</doi><tpages>5</tpages></addata></record> |
| fulltext | fulltext |
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| ispartof | Hybridoma, 1995-12, Vol.14 (6), p.587-591 |
| issn | 0272-457X 2168-7897 0272-457X |
| language | eng |
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| source | Mary Ann Liebert Online Subscription; MEDLINE |
| subjects | Animals Antibodies, immunoglobulins Antibodies, Monoclonal - biosynthesis Antibody Affinity Biological and medical sciences Biosensing Techniques Female Fundamental and applied biological sciences. Psychology Fundamental immunology Humans Hybridomas - chemistry Hybridomas - metabolism Membrane Glycoproteins - immunology Mice Mice, Inbred BALB C Milk, Human - immunology Molecular immunology Monoclonal antibodies Mucin-1 - immunology Precipitin Tests |
| title | Generation of murine monoclonal antihuman milk fat globule membrane antibodies using immunoprecipitation and BIAcore analyses |
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