Sorting polyclonal antibodies into functionally distinct fractions using peptide phage display: ‘a library on top of a library’
A general approach for sorting antibodies (Abs) to a restricted protein domain was developed using phage-displayed peptide libraries. The method is demonstrated by fractionating polyclonal antibodies (pAbs), raised against a short peptide derived from the extracellular, juxtamembrane region of fibro...
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| Veröffentlicht in: | Journal of immunological methods 2000-08, Vol.242 (1), p.183-191 |
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| container_title | Journal of immunological methods |
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| creator | Moshitch-Moshkovitz, S Heldman, Y Yayon, A Katchalski-Katzir, E |
| description | A general approach for sorting antibodies (Abs) to a restricted protein domain was developed using phage-displayed peptide libraries. The method is demonstrated by fractionating polyclonal antibodies (pAbs), raised against a short peptide derived from the extracellular, juxtamembrane region of fibroblast growth factor receptor 1 (FGFR1) into fractions with distinct chemical and biological characteristics. Screening two combinatorial peptide libraries, with the pAb, several sequences, homologous to different regions within the original peptide, were identified. Four of the corresponding peptides were synthesized and used as peptide-conjugated affinity columns for the fractionation of the pAbs. The fractions obtained were unique in their recognition patterns and in their capacity to immunoprecipitate and immunoblot, as well as to modulate the activity of FGFR1. This technique is, therefore, highly sufficient in separating pAbs to monospecific fractions and may also be used for fine mapping of different, even overlapping, sequences within a restricted peptide or protein domain. |
| doi_str_mv | 10.1016/S0022-1759(00)00247-7 |
| format | Article |
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The method is demonstrated by fractionating polyclonal antibodies (pAbs), raised against a short peptide derived from the extracellular, juxtamembrane region of fibroblast growth factor receptor 1 (FGFR1) into fractions with distinct chemical and biological characteristics. Screening two combinatorial peptide libraries, with the pAb, several sequences, homologous to different regions within the original peptide, were identified. Four of the corresponding peptides were synthesized and used as peptide-conjugated affinity columns for the fractionation of the pAbs. The fractions obtained were unique in their recognition patterns and in their capacity to immunoprecipitate and immunoblot, as well as to modulate the activity of FGFR1. 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Psychology ; Fundamental immunology ; Molecular immunology ; Molecular Sequence Data ; Peptide Library ; Peptide phage display ; Peptides - immunology ; Polyclonal antibodies ; Rabbits ; Receptor Protein-Tyrosine Kinases - immunology ; Receptor, Fibroblast Growth Factor, Type 1 ; Receptors, Fibroblast Growth Factor - immunology ; Techniques</subject><ispartof>Journal of immunological methods, 2000-08, Vol.242 (1), p.183-191</ispartof><rights>2000 Elsevier Science B.V.</rights><rights>2001 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c420t-de034ab8c587529f854347f8b7d359e2cd78065bd53ec7d16cbc2fc48133489f3</citedby><cites>FETCH-LOGICAL-c420t-de034ab8c587529f854347f8b7d359e2cd78065bd53ec7d16cbc2fc48133489f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/S0022-1759(00)00247-7$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=967131$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10986399$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Moshitch-Moshkovitz, S</creatorcontrib><creatorcontrib>Heldman, Y</creatorcontrib><creatorcontrib>Yayon, A</creatorcontrib><creatorcontrib>Katchalski-Katzir, E</creatorcontrib><title>Sorting polyclonal antibodies into functionally distinct fractions using peptide phage display: ‘a library on top of a library’</title><title>Journal of immunological methods</title><addtitle>J Immunol Methods</addtitle><description>A general approach for sorting antibodies (Abs) to a restricted protein domain was developed using phage-displayed peptide libraries. The method is demonstrated by fractionating polyclonal antibodies (pAbs), raised against a short peptide derived from the extracellular, juxtamembrane region of fibroblast growth factor receptor 1 (FGFR1) into fractions with distinct chemical and biological characteristics. Screening two combinatorial peptide libraries, with the pAb, several sequences, homologous to different regions within the original peptide, were identified. Four of the corresponding peptides were synthesized and used as peptide-conjugated affinity columns for the fractionation of the pAbs. The fractions obtained were unique in their recognition patterns and in their capacity to immunoprecipitate and immunoblot, as well as to modulate the activity of FGFR1. This technique is, therefore, highly sufficient in separating pAbs to monospecific fractions and may also be used for fine mapping of different, even overlapping, sequences within a restricted peptide or protein domain.</description><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Antibodies - immunology</subject><subject>Antibodies - isolation & purification</subject><subject>Biological and medical sciences</subject><subject>Cell Line</subject><subject>Chemical Fractionation</subject><subject>Chromatography, Affinity</subject><subject>Epitope library</subject><subject>Fibroblast growth factor receptor</subject><subject>fibroblast growth factor receptor 1</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fundamental immunology</subject><subject>Molecular immunology</subject><subject>Molecular Sequence Data</subject><subject>Peptide Library</subject><subject>Peptide phage display</subject><subject>Peptides - immunology</subject><subject>Polyclonal antibodies</subject><subject>Rabbits</subject><subject>Receptor Protein-Tyrosine Kinases - immunology</subject><subject>Receptor, Fibroblast Growth Factor, Type 1</subject><subject>Receptors, Fibroblast Growth Factor - immunology</subject><subject>Techniques</subject><issn>0022-1759</issn><issn>1872-7905</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc2KFDEQgIMo7rj6CEpAED20VjpJJ9mLyOIfLHhYPYd0kl4jmU6bpIW5LexLrK-3T2LPD4O3PYVUfVVJ1YfQcwJvCZDu3SVA2zZEcPUa4M1yYaIRD9CKSNE2QgF_iFZH5AQ9KeUXABDo4DE6IaBkR5VaoZvLlGsYr_CU4sbGNJqIzVhDn1zwBYexJjzMo61hm4ob7EJZeFvxkM0uWvBcdg38VIPzePpprvwWm6LZnOG761uDY-izyRucRlzThNOAj7G7679P0aPBxOKfHc5T9OPTx-_nX5qLb5-_nn-4aCxroTbOA2Wml5ZLwVs1SM4oE4PshaNc-dY6IaHjvePUW-FIZ3vbDpZJQimTaqCn6NW-75TT79mXqtehWB-jGX2aixZtyxVl7F6QiE5IwugC8j1ocyol-0FPOayXqTQBvdWkd5r01oEG0DtNWix1Lw4PzP3au_-q9l4W4OUBMMWauOx6tKEcOdUJQslCvd9Tftnan-CzLjb40XoXsrdVuxTu-cg_B76yXg</recordid><startdate>20000828</startdate><enddate>20000828</enddate><creator>Moshitch-Moshkovitz, S</creator><creator>Heldman, Y</creator><creator>Yayon, A</creator><creator>Katchalski-Katzir, E</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>7QO</scope><scope>7T5</scope><scope>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20000828</creationdate><title>Sorting polyclonal antibodies into functionally distinct fractions using peptide phage display: ‘a library on top of a library’</title><author>Moshitch-Moshkovitz, S ; Heldman, Y ; Yayon, A ; Katchalski-Katzir, E</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c420t-de034ab8c587529f854347f8b7d359e2cd78065bd53ec7d16cbc2fc48133489f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Antibodies - immunology</topic><topic>Antibodies - isolation & purification</topic><topic>Biological and medical sciences</topic><topic>Cell Line</topic><topic>Chemical Fractionation</topic><topic>Chromatography, Affinity</topic><topic>Epitope library</topic><topic>Fibroblast growth factor receptor</topic><topic>fibroblast growth factor receptor 1</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fundamental immunology</topic><topic>Molecular immunology</topic><topic>Molecular Sequence Data</topic><topic>Peptide Library</topic><topic>Peptide phage display</topic><topic>Peptides - immunology</topic><topic>Polyclonal antibodies</topic><topic>Rabbits</topic><topic>Receptor Protein-Tyrosine Kinases - immunology</topic><topic>Receptor, Fibroblast Growth Factor, Type 1</topic><topic>Receptors, Fibroblast Growth Factor - immunology</topic><topic>Techniques</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Moshitch-Moshkovitz, S</creatorcontrib><creatorcontrib>Heldman, Y</creatorcontrib><creatorcontrib>Yayon, A</creatorcontrib><creatorcontrib>Katchalski-Katzir, E</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>Biotechnology Research Abstracts</collection><collection>Immunology Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of immunological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Moshitch-Moshkovitz, S</au><au>Heldman, Y</au><au>Yayon, A</au><au>Katchalski-Katzir, E</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Sorting polyclonal antibodies into functionally distinct fractions using peptide phage display: ‘a library on top of a library’</atitle><jtitle>Journal of immunological methods</jtitle><addtitle>J Immunol Methods</addtitle><date>2000-08-28</date><risdate>2000</risdate><volume>242</volume><issue>1</issue><spage>183</spage><epage>191</epage><pages>183-191</pages><issn>0022-1759</issn><eissn>1872-7905</eissn><coden>JIMMBG</coden><abstract>A general approach for sorting antibodies (Abs) to a restricted protein domain was developed using phage-displayed peptide libraries. The method is demonstrated by fractionating polyclonal antibodies (pAbs), raised against a short peptide derived from the extracellular, juxtamembrane region of fibroblast growth factor receptor 1 (FGFR1) into fractions with distinct chemical and biological characteristics. Screening two combinatorial peptide libraries, with the pAb, several sequences, homologous to different regions within the original peptide, were identified. Four of the corresponding peptides were synthesized and used as peptide-conjugated affinity columns for the fractionation of the pAbs. The fractions obtained were unique in their recognition patterns and in their capacity to immunoprecipitate and immunoblot, as well as to modulate the activity of FGFR1. 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| subjects | Amino Acid Sequence Animals Antibodies - immunology Antibodies - isolation & purification Biological and medical sciences Cell Line Chemical Fractionation Chromatography, Affinity Epitope library Fibroblast growth factor receptor fibroblast growth factor receptor 1 Fundamental and applied biological sciences. Psychology Fundamental immunology Molecular immunology Molecular Sequence Data Peptide Library Peptide phage display Peptides - immunology Polyclonal antibodies Rabbits Receptor Protein-Tyrosine Kinases - immunology Receptor, Fibroblast Growth Factor, Type 1 Receptors, Fibroblast Growth Factor - immunology Techniques |
| title | Sorting polyclonal antibodies into functionally distinct fractions using peptide phage display: ‘a library on top of a library’ |
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