Recombinant antibody fragments that detect enoyl acyl carrier protein reductase in Brassica napus
Purified Brassica napus enoyl acyl carrier protein reductase (ENR) was used to select specific antibodies from a library of antibody fragments, single‐chain Fv (scFv), displayed on filamentous phage. Analysis of the selected clones by BstNl fingerprinting and nucleotide sequencing showed that the sc...
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| Veröffentlicht in: | Lipids 1997-08, Vol.32 (8), p.805-809 |
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| creator | Ziegler, A. (Scottish Crop Research Institute, Invergowrie, Dundee, UK.) Macintosh, S.M Torrance, L Simon, W Slabas, A.R |
| description | Purified Brassica napus enoyl acyl carrier protein reductase (ENR) was used to select specific antibodies from a library of antibody fragments, single‐chain Fv (scFv), displayed on filamentous phage. Analysis of the selected clones by BstNl fingerprinting and nucleotide sequencing showed that the scFv were derived from three different human VH germline genes. The binding specificities were confirmed by Western blots and ELISA. The scFv preparations reacted with B. napus ENR, but not with β‐keto reductase, nor enoyl reductase from Escherichia coli. Analysis of fragments generated by CNBr treatment indicates that the scFv 3.13 recognizes an epitope located within the n‐terminal 80 amino acids of the enzyme molecule. The scFv were used to detect ENR directly in extracts of B. napus seeds. |
| doi_str_mv | 10.1007/s11745-997-0103-3 |
| format | Article |
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(Scottish Crop Research Institute, Invergowrie, Dundee, UK.) ; Macintosh, S.M ; Torrance, L ; Simon, W ; Slabas, A.R</creator><creatorcontrib>Ziegler, A. (Scottish Crop Research Institute, Invergowrie, Dundee, UK.) ; Macintosh, S.M ; Torrance, L ; Simon, W ; Slabas, A.R</creatorcontrib><description>Purified Brassica napus enoyl acyl carrier protein reductase (ENR) was used to select specific antibodies from a library of antibody fragments, single‐chain Fv (scFv), displayed on filamentous phage. Analysis of the selected clones by BstNl fingerprinting and nucleotide sequencing showed that the scFv were derived from three different human VH germline genes. The binding specificities were confirmed by Western blots and ELISA. The scFv preparations reacted with B. napus ENR, but not with β‐keto reductase, nor enoyl reductase from Escherichia coli. Analysis of fragments generated by CNBr treatment indicates that the scFv 3.13 recognizes an epitope located within the n‐terminal 80 amino acids of the enzyme molecule. The scFv were used to detect ENR directly in extracts of B. napus seeds.</description><identifier>ISSN: 0024-4201</identifier><identifier>EISSN: 1558-9307</identifier><identifier>DOI: 10.1007/s11745-997-0103-3</identifier><identifier>PMID: 9270971</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer‐Verlag</publisher><subject>Acyl Carrier Protein - metabolism ; Amino Acid Sequence ; Amino acids ; Bacteria ; Bacteriophages - genetics ; Blotting, Western ; Brassica - enzymology ; BRASSICA NAPUS ; Cloning, Molecular ; E coli ; Enoyl-(Acyl-Carrier-Protein) Reductase (NADH) ; Enzyme-Linked Immunosorbent Assay ; Epitope Mapping ; Escherichia coli - enzymology ; Escherichia coli - genetics ; Fatty Acid Synthases - analysis ; Fatty Acid Synthases - immunology ; Humans ; Immunoglobulin Fragments - chemistry ; Immunoglobulin Fragments - genetics ; Immunoglobulin Fragments - immunology ; Immunoglobulin Fragments - metabolism ; Molecular Sequence Data ; Oxidoreductases - analysis ; Oxidoreductases - immunology ; Peptide Library ; Recombinant Proteins - chemistry ; Recombinant Proteins - immunology ; Recombinant Proteins - metabolism ; Seeds ; Seeds - enzymology ; Sequence Alignment</subject><ispartof>Lipids, 1997-08, Vol.32 (8), p.805-809</ispartof><rights>1997 American Oil Chemists' Society (AOCS)</rights><rights>AOCS Press 1997</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3905-7fe5996ef46392542ec5626fa2588334e765eaff89587caa701ffedc3b8cc16c3</citedby><cites>FETCH-LOGICAL-c3905-7fe5996ef46392542ec5626fa2588334e765eaff89587caa701ffedc3b8cc16c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1007%2Fs11745-997-0103-3$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1007%2Fs11745-997-0103-3$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9270971$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ziegler, A. (Scottish Crop Research Institute, Invergowrie, Dundee, UK.)</creatorcontrib><creatorcontrib>Macintosh, S.M</creatorcontrib><creatorcontrib>Torrance, L</creatorcontrib><creatorcontrib>Simon, W</creatorcontrib><creatorcontrib>Slabas, A.R</creatorcontrib><title>Recombinant antibody fragments that detect enoyl acyl carrier protein reductase in Brassica napus</title><title>Lipids</title><addtitle>Lipids</addtitle><description>Purified Brassica napus enoyl acyl carrier protein reductase (ENR) was used to select specific antibodies from a library of antibody fragments, single‐chain Fv (scFv), displayed on filamentous phage. Analysis of the selected clones by BstNl fingerprinting and nucleotide sequencing showed that the scFv were derived from three different human VH germline genes. The binding specificities were confirmed by Western blots and ELISA. The scFv preparations reacted with B. napus ENR, but not with β‐keto reductase, nor enoyl reductase from Escherichia coli. Analysis of fragments generated by CNBr treatment indicates that the scFv 3.13 recognizes an epitope located within the n‐terminal 80 amino acids of the enzyme molecule. The scFv were used to detect ENR directly in extracts of B. napus seeds.</description><subject>Acyl Carrier Protein - metabolism</subject><subject>Amino Acid Sequence</subject><subject>Amino acids</subject><subject>Bacteria</subject><subject>Bacteriophages - genetics</subject><subject>Blotting, Western</subject><subject>Brassica - enzymology</subject><subject>BRASSICA NAPUS</subject><subject>Cloning, Molecular</subject><subject>E coli</subject><subject>Enoyl-(Acyl-Carrier-Protein) Reductase (NADH)</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Epitope Mapping</subject><subject>Escherichia coli - enzymology</subject><subject>Escherichia coli - genetics</subject><subject>Fatty Acid Synthases - analysis</subject><subject>Fatty Acid Synthases - immunology</subject><subject>Humans</subject><subject>Immunoglobulin Fragments - chemistry</subject><subject>Immunoglobulin Fragments - genetics</subject><subject>Immunoglobulin Fragments - immunology</subject><subject>Immunoglobulin Fragments - metabolism</subject><subject>Molecular Sequence Data</subject><subject>Oxidoreductases - analysis</subject><subject>Oxidoreductases - immunology</subject><subject>Peptide Library</subject><subject>Recombinant Proteins - chemistry</subject><subject>Recombinant Proteins - immunology</subject><subject>Recombinant Proteins - metabolism</subject><subject>Seeds</subject><subject>Seeds - enzymology</subject><subject>Sequence Alignment</subject><issn>0024-4201</issn><issn>1558-9307</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1997</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqFkE2LFDEQhoMo6zj6AxSE4MFbu0kn6SRHd_1aGNhF3XOoSVfWLP0xJmmW-fdm6cGDFw9VRVHv-1I8hLzm7ANnTJ9nzrVUjbW6YZyJRjwhG66Uaaxg-inZMNbKRraMPycvcr6vK5dWnZEz22pmNd8Q-I5-HvdxgqnQWnE_90caEtyNOJVMyy8otMeCvlCc5uNAwdfmIaWIiR7SXDBONGG_-AIZaV0uEuQcPdAJDkt-SZ4FGDK-Os0tuf3y-eflt2Z3_fXq8uOu8cIy1eiAytoOg-yEbZVs0auu7QK0yhghJOpOIYRgrDLaA2jGQ8Dei73xnndebMn7Nbf-9HvBXNwYs8dhgAnnJTttW8lFpbQl7_4R3s9LmupvzhjDhRTMVhFfRT7NOScM7pDiCOnoOHOP7N3K3lX27pG9E9Xz9hS87Efs_zpOsOtdr_eHOODx_4Fud3XziRmmqvPN6gwwO7hLMbvbH1YLrbUVfwDBtJh8</recordid><startdate>199708</startdate><enddate>199708</enddate><creator>Ziegler, A. (Scottish Crop Research Institute, Invergowrie, Dundee, UK.)</creator><creator>Macintosh, S.M</creator><creator>Torrance, L</creator><creator>Simon, W</creator><creator>Slabas, A.R</creator><general>Springer‐Verlag</general><general>Springer Nature B.V</general><scope>FBQ</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>3V.</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7T7</scope><scope>7TK</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>BKSAR</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PCBAR</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>7X8</scope></search><sort><creationdate>199708</creationdate><title>Recombinant antibody fragments that detect enoyl acyl carrier protein reductase in Brassica napus</title><author>Ziegler, A. 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(Scottish Crop Research Institute, Invergowrie, Dundee, UK.)</creatorcontrib><creatorcontrib>Macintosh, S.M</creatorcontrib><creatorcontrib>Torrance, L</creatorcontrib><creatorcontrib>Simon, W</creatorcontrib><creatorcontrib>Slabas, A.R</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Neurosciences Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>Earth, Atmospheric & Aquatic Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Earth, Atmospheric & Aquatic Science Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>MEDLINE - Academic</collection><jtitle>Lipids</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ziegler, A. (Scottish Crop Research Institute, Invergowrie, Dundee, UK.)</au><au>Macintosh, S.M</au><au>Torrance, L</au><au>Simon, W</au><au>Slabas, A.R</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Recombinant antibody fragments that detect enoyl acyl carrier protein reductase in Brassica napus</atitle><jtitle>Lipids</jtitle><addtitle>Lipids</addtitle><date>1997-08</date><risdate>1997</risdate><volume>32</volume><issue>8</issue><spage>805</spage><epage>809</epage><pages>805-809</pages><issn>0024-4201</issn><eissn>1558-9307</eissn><abstract>Purified Brassica napus enoyl acyl carrier protein reductase (ENR) was used to select specific antibodies from a library of antibody fragments, single‐chain Fv (scFv), displayed on filamentous phage. Analysis of the selected clones by BstNl fingerprinting and nucleotide sequencing showed that the scFv were derived from three different human VH germline genes. The binding specificities were confirmed by Western blots and ELISA. The scFv preparations reacted with B. napus ENR, but not with β‐keto reductase, nor enoyl reductase from Escherichia coli. Analysis of fragments generated by CNBr treatment indicates that the scFv 3.13 recognizes an epitope located within the n‐terminal 80 amino acids of the enzyme molecule. The scFv were used to detect ENR directly in extracts of B. napus seeds.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer‐Verlag</pub><pmid>9270971</pmid><doi>10.1007/s11745-997-0103-3</doi><tpages>5</tpages></addata></record> |
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| ispartof | Lipids, 1997-08, Vol.32 (8), p.805-809 |
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| language | eng |
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| subjects | Acyl Carrier Protein - metabolism Amino Acid Sequence Amino acids Bacteria Bacteriophages - genetics Blotting, Western Brassica - enzymology BRASSICA NAPUS Cloning, Molecular E coli Enoyl-(Acyl-Carrier-Protein) Reductase (NADH) Enzyme-Linked Immunosorbent Assay Epitope Mapping Escherichia coli - enzymology Escherichia coli - genetics Fatty Acid Synthases - analysis Fatty Acid Synthases - immunology Humans Immunoglobulin Fragments - chemistry Immunoglobulin Fragments - genetics Immunoglobulin Fragments - immunology Immunoglobulin Fragments - metabolism Molecular Sequence Data Oxidoreductases - analysis Oxidoreductases - immunology Peptide Library Recombinant Proteins - chemistry Recombinant Proteins - immunology Recombinant Proteins - metabolism Seeds Seeds - enzymology Sequence Alignment |
| title | Recombinant antibody fragments that detect enoyl acyl carrier protein reductase in Brassica napus |
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