Comparative interaction kinetics of two recombinant fabs and of the corresponding antibodies directed to the coat protein of tobacco mosaic virus
Two recombinant Fab fragments, 57P and 174P, recognizing peptide 134–146 of the coat protein of tobacco mosaic virus have been cloned, sequenced and expressed in Escherichia coli. They differ by 15 amino acid changes in the sequence of their variable region. The interaction kinetics of the Fabs with...
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| Veröffentlicht in: | Journal of molecular recognition 1996-01, Vol.9 (1), p.39-51 |
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| creator | Chatellier, Jean Rauffer-Bruyère, Nathalie Van Regenmortel, Marc H. V. Altschuh, Danièle Weiss, Etienne |
| description | Two recombinant Fab fragments, 57P and 174P, recognizing peptide 134–146 of the coat protein of tobacco mosaic virus have been cloned, sequenced and expressed in Escherichia coli. They differ by 15 amino acid changes in the sequence of their variable region. The interaction kinetics of the Fabs with the wild‐type and four mutant peptides have been compared using a BIAcoreTM biosensor instrument. The recombinant Fab 174P had the same reactivity as the Fab fragment obtained by enzymatic cleavage of monoclonal antibody 174P. The two recombinant Fabs recognized the various peptides in the same ranking order but Fab 174P consistently dissociated somewhat faster from the peptides compared to Fab 57P. The two whole antibodies showed the same relative differences in reactivity as the two recombinant Fabs. The location of amino acid changes was visualized on a model structure of the Fab. Differences in dissociation rates of the two antibodies are most likely due to changes located at the periphery of the antigen‐combining site and/or at the interface between the light and heavy chain domains. Our results demonstrate the feasibility of detecting very small differences in binding affinity by the biosensor technology, which is a prerequisite for assessing the functional effect of limited structural changes. |
| doi_str_mv | 10.1002/(SICI)1099-1352(199601)9:1<39::AID-JMR239>3.0.CO;2-V |
| format | Article |
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V. ; Altschuh, Danièle ; Weiss, Etienne</creator><creatorcontrib>Chatellier, Jean ; Rauffer-Bruyère, Nathalie ; Van Regenmortel, Marc H. V. ; Altschuh, Danièle ; Weiss, Etienne</creatorcontrib><description>Two recombinant Fab fragments, 57P and 174P, recognizing peptide 134–146 of the coat protein of tobacco mosaic virus have been cloned, sequenced and expressed in Escherichia coli. They differ by 15 amino acid changes in the sequence of their variable region. The interaction kinetics of the Fabs with the wild‐type and four mutant peptides have been compared using a BIAcoreTM biosensor instrument. The recombinant Fab 174P had the same reactivity as the Fab fragment obtained by enzymatic cleavage of monoclonal antibody 174P. The two recombinant Fabs recognized the various peptides in the same ranking order but Fab 174P consistently dissociated somewhat faster from the peptides compared to Fab 57P. The two whole antibodies showed the same relative differences in reactivity as the two recombinant Fabs. The location of amino acid changes was visualized on a model structure of the Fab. Differences in dissociation rates of the two antibodies are most likely due to changes located at the periphery of the antigen‐combining site and/or at the interface between the light and heavy chain domains. Our results demonstrate the feasibility of detecting very small differences in binding affinity by the biosensor technology, which is a prerequisite for assessing the functional effect of limited structural changes.</description><identifier>ISSN: 0952-3499</identifier><identifier>EISSN: 1099-1352</identifier><identifier>DOI: 10.1002/(SICI)1099-1352(199601)9:1<39::AID-JMR239>3.0.CO;2-V</identifier><identifier>PMID: 8723318</identifier><language>eng</language><publisher>Chichester, UK: John Wiley & Sons, Ltd</publisher><subject>Amino Acid Sequence ; Antibodies, Monoclonal ; Antibodies, Monoclonal - immunology ; Antibodies, Monoclonal - metabolism ; Antibodies, Viral ; Antibodies, Viral - chemistry ; Antibodies, Viral - immunology ; Antibodies, Viral - metabolism ; Antigen Presentation ; BIAcoreTM ; Biochemistry, Molecular Biology ; Biosensing Techniques ; Capsid ; Capsid - chemistry ; Capsid - immunology ; Capsid - metabolism ; cloning strategy ; Cloning, Molecular ; DNA Primers ; Escherichia coli ; Escherichia coli - genetics ; filter screening ; Immunoglobulin Fab Fragments ; Immunoglobulin Fab Fragments - chemistry ; Immunoglobulin Fab Fragments - immunology ; Immunoglobulin Fab Fragments - metabolism ; Immunoglobulin Variable Region ; Immunoglobulin Variable Region - chemistry ; kinetic rate constants ; Kinetics ; Life Sciences ; Models, Molecular ; Molecular Sequence Data ; Mutation ; Peptide Fragments ; Peptide Fragments - chemistry ; Peptide Fragments - metabolism ; Peptide Fragments - pharmacology ; peptide-antibody interaction ; recombinant Fab ; Recombinant Proteins ; Recombinant Proteins - immunology ; Recombinant Proteins - metabolism ; Tobacco Mosaic Virus ; Tobacco Mosaic Virus - immunology ; tobacco mosaic virus protein</subject><ispartof>Journal of molecular recognition, 1996-01, Vol.9 (1), p.39-51</ispartof><rights>Copyright © 1996 John Wiley & Sons Ltd.</rights><rights>Distributed under a Creative Commons Attribution 4.0 International License</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c4349-72289f944ea45d8c55937e75c3384f8afefef79553747738bf4d600c2c12d2683</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2F%28SICI%291099-1352%28199601%299%3A1%3C39%3A%3AAID-JMR239%3E3.0.CO%3B2-V$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2F%28SICI%291099-1352%28199601%299%3A1%3C39%3A%3AAID-JMR239%3E3.0.CO%3B2-V$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>230,314,776,780,881,1411,4010,27900,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8723318$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://hal.science/hal-00265268$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Chatellier, Jean</creatorcontrib><creatorcontrib>Rauffer-Bruyère, Nathalie</creatorcontrib><creatorcontrib>Van Regenmortel, Marc H. V.</creatorcontrib><creatorcontrib>Altschuh, Danièle</creatorcontrib><creatorcontrib>Weiss, Etienne</creatorcontrib><title>Comparative interaction kinetics of two recombinant fabs and of the corresponding antibodies directed to the coat protein of tobacco mosaic virus</title><title>Journal of molecular recognition</title><addtitle>J. Mol. Recognit</addtitle><description>Two recombinant Fab fragments, 57P and 174P, recognizing peptide 134–146 of the coat protein of tobacco mosaic virus have been cloned, sequenced and expressed in Escherichia coli. They differ by 15 amino acid changes in the sequence of their variable region. The interaction kinetics of the Fabs with the wild‐type and four mutant peptides have been compared using a BIAcoreTM biosensor instrument. The recombinant Fab 174P had the same reactivity as the Fab fragment obtained by enzymatic cleavage of monoclonal antibody 174P. The two recombinant Fabs recognized the various peptides in the same ranking order but Fab 174P consistently dissociated somewhat faster from the peptides compared to Fab 57P. The two whole antibodies showed the same relative differences in reactivity as the two recombinant Fabs. The location of amino acid changes was visualized on a model structure of the Fab. Differences in dissociation rates of the two antibodies are most likely due to changes located at the periphery of the antigen‐combining site and/or at the interface between the light and heavy chain domains. Our results demonstrate the feasibility of detecting very small differences in binding affinity by the biosensor technology, which is a prerequisite for assessing the functional effect of limited structural changes.</description><subject>Amino Acid Sequence</subject><subject>Antibodies, Monoclonal</subject><subject>Antibodies, Monoclonal - immunology</subject><subject>Antibodies, Monoclonal - metabolism</subject><subject>Antibodies, Viral</subject><subject>Antibodies, Viral - chemistry</subject><subject>Antibodies, Viral - immunology</subject><subject>Antibodies, Viral - metabolism</subject><subject>Antigen Presentation</subject><subject>BIAcoreTM</subject><subject>Biochemistry, Molecular Biology</subject><subject>Biosensing Techniques</subject><subject>Capsid</subject><subject>Capsid - chemistry</subject><subject>Capsid - immunology</subject><subject>Capsid - metabolism</subject><subject>cloning strategy</subject><subject>Cloning, Molecular</subject><subject>DNA Primers</subject><subject>Escherichia coli</subject><subject>Escherichia coli - genetics</subject><subject>filter screening</subject><subject>Immunoglobulin Fab Fragments</subject><subject>Immunoglobulin Fab Fragments - chemistry</subject><subject>Immunoglobulin Fab Fragments - immunology</subject><subject>Immunoglobulin Fab Fragments - metabolism</subject><subject>Immunoglobulin Variable Region</subject><subject>Immunoglobulin Variable Region - chemistry</subject><subject>kinetic rate constants</subject><subject>Kinetics</subject><subject>Life Sciences</subject><subject>Models, Molecular</subject><subject>Molecular Sequence Data</subject><subject>Mutation</subject><subject>Peptide Fragments</subject><subject>Peptide Fragments - chemistry</subject><subject>Peptide Fragments - metabolism</subject><subject>Peptide Fragments - pharmacology</subject><subject>peptide-antibody interaction</subject><subject>recombinant Fab</subject><subject>Recombinant Proteins</subject><subject>Recombinant Proteins - immunology</subject><subject>Recombinant Proteins - metabolism</subject><subject>Tobacco Mosaic Virus</subject><subject>Tobacco Mosaic Virus - immunology</subject><subject>tobacco mosaic virus protein</subject><issn>0952-3499</issn><issn>1099-1352</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kV1v0zAYhSMEGmXwE5B8hdaLFH8kjV0mpCrQrqisfIxy-cp1HOatiYvtduxn8I9xl6o3IOQLyz7nPcfWkyTnBA8IxvT12ddZOesTLERKWE7PiBBDTPpiRM6ZGI3Gs3fph49fKBNv2QAPysUbmi4fJb3jwOOkh0VOU5YJ8TR55v0NxlHL8UlywgvKGOG95Hdpm410MpidRqYN2kkVjG3RrWl1MMojW6NwZ5HTyjYr08o2oFquPJJt9aBda6Ssc9pvbFuZ9kcUglnZymiPKhPHgq5QsAejDGjjbNCmfRi2K6mURY310ii0M27rnydParn2-sVhP02-Td5flRfpfDGdleN5qrL4o7SglItaZJmWWV5xleeCFbrIFWM8q7msdVyFyHNWZEXB-KrOqiHGiipCKzrk7DTpd7nXcg0bZxrp7sFKAxfjOezvIoNhHp07Er2vOm98-8-t9gEa45Ver2Wr7dZDwYlgnGfReNUZlbPeO10fkwmGPVWAPVXYQ4I9JOioQjwBEwCRKnRUgQGGcgEUljH25aF_u2p0dQw9YIz6stPvzFrf_9X538p_Nh5uYnDaBRsf9K9jsHS3MCxYkcP3yylM-OXk8ySbwif2B782zfM</recordid><startdate>199601</startdate><enddate>199601</enddate><creator>Chatellier, Jean</creator><creator>Rauffer-Bruyère, Nathalie</creator><creator>Van Regenmortel, Marc H. V.</creator><creator>Altschuh, Danièle</creator><creator>Weiss, Etienne</creator><general>John Wiley & Sons, Ltd</general><general>Wiley</general><scope>BSCLL</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>1XC</scope></search><sort><creationdate>199601</creationdate><title>Comparative interaction kinetics of two recombinant fabs and of the corresponding antibodies directed to the coat protein of tobacco mosaic virus</title><author>Chatellier, Jean ; Rauffer-Bruyère, Nathalie ; Van Regenmortel, Marc H. V. ; Altschuh, Danièle ; Weiss, Etienne</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4349-72289f944ea45d8c55937e75c3384f8afefef79553747738bf4d600c2c12d2683</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Amino Acid Sequence</topic><topic>Antibodies, Monoclonal</topic><topic>Antibodies, Monoclonal - immunology</topic><topic>Antibodies, Monoclonal - metabolism</topic><topic>Antibodies, Viral</topic><topic>Antibodies, Viral - chemistry</topic><topic>Antibodies, Viral - immunology</topic><topic>Antibodies, Viral - metabolism</topic><topic>Antigen Presentation</topic><topic>BIAcoreTM</topic><topic>Biochemistry, Molecular Biology</topic><topic>Biosensing Techniques</topic><topic>Capsid</topic><topic>Capsid - chemistry</topic><topic>Capsid - immunology</topic><topic>Capsid - metabolism</topic><topic>cloning strategy</topic><topic>Cloning, Molecular</topic><topic>DNA Primers</topic><topic>Escherichia coli</topic><topic>Escherichia coli - genetics</topic><topic>filter screening</topic><topic>Immunoglobulin Fab Fragments</topic><topic>Immunoglobulin Fab Fragments - chemistry</topic><topic>Immunoglobulin Fab Fragments - immunology</topic><topic>Immunoglobulin Fab Fragments - metabolism</topic><topic>Immunoglobulin Variable Region</topic><topic>Immunoglobulin Variable Region - chemistry</topic><topic>kinetic rate constants</topic><topic>Kinetics</topic><topic>Life Sciences</topic><topic>Models, Molecular</topic><topic>Molecular Sequence Data</topic><topic>Mutation</topic><topic>Peptide Fragments</topic><topic>Peptide Fragments - chemistry</topic><topic>Peptide Fragments - metabolism</topic><topic>Peptide Fragments - pharmacology</topic><topic>peptide-antibody interaction</topic><topic>recombinant Fab</topic><topic>Recombinant Proteins</topic><topic>Recombinant Proteins - immunology</topic><topic>Recombinant Proteins - metabolism</topic><topic>Tobacco Mosaic Virus</topic><topic>Tobacco Mosaic Virus - immunology</topic><topic>tobacco mosaic virus protein</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chatellier, Jean</creatorcontrib><creatorcontrib>Rauffer-Bruyère, Nathalie</creatorcontrib><creatorcontrib>Van Regenmortel, Marc H. V.</creatorcontrib><creatorcontrib>Altschuh, Danièle</creatorcontrib><creatorcontrib>Weiss, Etienne</creatorcontrib><collection>Istex</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>Hyper Article en Ligne (HAL)</collection><jtitle>Journal of molecular recognition</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chatellier, Jean</au><au>Rauffer-Bruyère, Nathalie</au><au>Van Regenmortel, Marc H. V.</au><au>Altschuh, Danièle</au><au>Weiss, Etienne</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Comparative interaction kinetics of two recombinant fabs and of the corresponding antibodies directed to the coat protein of tobacco mosaic virus</atitle><jtitle>Journal of molecular recognition</jtitle><addtitle>J. Mol. Recognit</addtitle><date>1996-01</date><risdate>1996</risdate><volume>9</volume><issue>1</issue><spage>39</spage><epage>51</epage><pages>39-51</pages><issn>0952-3499</issn><eissn>1099-1352</eissn><abstract>Two recombinant Fab fragments, 57P and 174P, recognizing peptide 134–146 of the coat protein of tobacco mosaic virus have been cloned, sequenced and expressed in Escherichia coli. They differ by 15 amino acid changes in the sequence of their variable region. The interaction kinetics of the Fabs with the wild‐type and four mutant peptides have been compared using a BIAcoreTM biosensor instrument. The recombinant Fab 174P had the same reactivity as the Fab fragment obtained by enzymatic cleavage of monoclonal antibody 174P. The two recombinant Fabs recognized the various peptides in the same ranking order but Fab 174P consistently dissociated somewhat faster from the peptides compared to Fab 57P. The two whole antibodies showed the same relative differences in reactivity as the two recombinant Fabs. The location of amino acid changes was visualized on a model structure of the Fab. Differences in dissociation rates of the two antibodies are most likely due to changes located at the periphery of the antigen‐combining site and/or at the interface between the light and heavy chain domains. Our results demonstrate the feasibility of detecting very small differences in binding affinity by the biosensor technology, which is a prerequisite for assessing the functional effect of limited structural changes.</abstract><cop>Chichester, UK</cop><pub>John Wiley & Sons, Ltd</pub><pmid>8723318</pmid><doi>10.1002/(SICI)1099-1352(199601)9:1<39::AID-JMR239>3.0.CO;2-V</doi><tpages>13</tpages></addata></record> |
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| identifier | ISSN: 0952-3499 |
| ispartof | Journal of molecular recognition, 1996-01, Vol.9 (1), p.39-51 |
| issn | 0952-3499 1099-1352 |
| language | eng |
| recordid | cdi_hal_primary_oai_HAL_hal_00265268v1 |
| source | MEDLINE; Wiley Online Library Journals Frontfile Complete |
| subjects | Amino Acid Sequence Antibodies, Monoclonal Antibodies, Monoclonal - immunology Antibodies, Monoclonal - metabolism Antibodies, Viral Antibodies, Viral - chemistry Antibodies, Viral - immunology Antibodies, Viral - metabolism Antigen Presentation BIAcoreTM Biochemistry, Molecular Biology Biosensing Techniques Capsid Capsid - chemistry Capsid - immunology Capsid - metabolism cloning strategy Cloning, Molecular DNA Primers Escherichia coli Escherichia coli - genetics filter screening Immunoglobulin Fab Fragments Immunoglobulin Fab Fragments - chemistry Immunoglobulin Fab Fragments - immunology Immunoglobulin Fab Fragments - metabolism Immunoglobulin Variable Region Immunoglobulin Variable Region - chemistry kinetic rate constants Kinetics Life Sciences Models, Molecular Molecular Sequence Data Mutation Peptide Fragments Peptide Fragments - chemistry Peptide Fragments - metabolism Peptide Fragments - pharmacology peptide-antibody interaction recombinant Fab Recombinant Proteins Recombinant Proteins - immunology Recombinant Proteins - metabolism Tobacco Mosaic Virus Tobacco Mosaic Virus - immunology tobacco mosaic virus protein |
| title | Comparative interaction kinetics of two recombinant fabs and of the corresponding antibodies directed to the coat protein of tobacco mosaic virus |
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