Antibody arrays for high-throughput screening of antibody-antigen interactions
We have developed a novel technique for high-throughput screening of recombinant antibodies, based on the creation of antibody arrays. Our method uses robotic picking and high-density gridding of bacteria containing antibody genes followed by filter-based enzyme-linked immunosorbent assay (ELISA) sc...
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| Veröffentlicht in: | Nature biotechnology 2000-09, Vol.18 (9), p.989-994 |
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| creator | de Wildt, Ruud M. T Tomlinson, Ian M Mundy, Chris R Gorick, Barbara D |
| description | We have developed a novel technique for high-throughput screening of recombinant antibodies, based on the creation of antibody arrays. Our method uses robotic picking and high-density gridding of bacteria containing antibody genes followed by filter-based enzyme-linked immunosorbent assay (ELISA) screening to identify clones that express binding antibody fragments. By eliminating the need for liquid handling, we can thereby screen up to 18,342 different antibody clones at a time and, because the clones are arrayed from master stocks, the same antibodies can be double spotted and screened simultaneously against 15 different antigens. We have used our technique in several different applications, including isolating antibodies against impure proteins and complex antigens, where several rounds of phage display often fail. Our results indicate that antibody arrays can be used to identify differentially expressed proteins. |
| doi_str_mv | 10.1038/79494 |
| format | Article |
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T ; Tomlinson, Ian M ; Mundy, Chris R ; Gorick, Barbara D</creator><creatorcontrib>de Wildt, Ruud M. T ; Tomlinson, Ian M ; Mundy, Chris R ; Gorick, Barbara D</creatorcontrib><description>We have developed a novel technique for high-throughput screening of recombinant antibodies, based on the creation of antibody arrays. Our method uses robotic picking and high-density gridding of bacteria containing antibody genes followed by filter-based enzyme-linked immunosorbent assay (ELISA) screening to identify clones that express binding antibody fragments. By eliminating the need for liquid handling, we can thereby screen up to 18,342 different antibody clones at a time and, because the clones are arrayed from master stocks, the same antibodies can be double spotted and screened simultaneously against 15 different antigens. We have used our technique in several different applications, including isolating antibodies against impure proteins and complex antigens, where several rounds of phage display often fail. Our results indicate that antibody arrays can be used to identify differentially expressed proteins.</description><identifier>ISSN: 1087-0156</identifier><identifier>EISSN: 1546-1696</identifier><identifier>DOI: 10.1038/79494</identifier><identifier>PMID: 10973222</identifier><identifier>CODEN: NABIF9</identifier><language>eng</language><publisher>New York, NY: Nature</publisher><subject>Amino Acid Sequence ; Antibodies ; Antibodies - chemistry ; antibody arrays ; Antigen-Antibody Reactions ; Antigens ; Arrays ; Bacteria - chemistry ; Bacteria - genetics ; Biochemistry - methods ; Biological and medical sciences ; Biosensing Techniques - methods ; Biotechnology ; Blotting, Western ; Cloning ; Enzyme-Linked Immunosorbent Assay ; Enzymes ; Fundamental and applied biological sciences. Psychology ; HeLa Cells ; high-throughput screening ; Humans ; Ligands ; Methods. Procedures. Technologies ; Molecular Probe Techniques ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Others ; Peptide Library ; Protein Conformation ; Proteins ; Proteins - chemistry ; Proteins - metabolism ; Recombinant Proteins - chemistry ; Robotics ; Serum Albumin - chemistry ; Serum Albumin, Bovine - chemistry ; Various methods and equipments</subject><ispartof>Nature biotechnology, 2000-09, Vol.18 (9), p.989-994</ispartof><rights>2000 INIST-CNRS</rights><rights>COPYRIGHT 2000 Nature Publishing Group</rights><rights>Copyright Nature Publishing Group Sep 2000</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c628t-3e18cd0aebfa24ddd2e6b540989d48d5c0a806c688e0e9328aee2f515f6fe33</citedby><cites>FETCH-LOGICAL-c628t-3e18cd0aebfa24ddd2e6b540989d48d5c0a806c688e0e9328aee2f515f6fe33</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,782,786,2729,27931,27932</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=1504427$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10973222$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>de Wildt, Ruud M. T</creatorcontrib><creatorcontrib>Tomlinson, Ian M</creatorcontrib><creatorcontrib>Mundy, Chris R</creatorcontrib><creatorcontrib>Gorick, Barbara D</creatorcontrib><title>Antibody arrays for high-throughput screening of antibody-antigen interactions</title><title>Nature biotechnology</title><addtitle>Nat Biotechnol</addtitle><description>We have developed a novel technique for high-throughput screening of recombinant antibodies, based on the creation of antibody arrays. Our method uses robotic picking and high-density gridding of bacteria containing antibody genes followed by filter-based enzyme-linked immunosorbent assay (ELISA) screening to identify clones that express binding antibody fragments. By eliminating the need for liquid handling, we can thereby screen up to 18,342 different antibody clones at a time and, because the clones are arrayed from master stocks, the same antibodies can be double spotted and screened simultaneously against 15 different antigens. We have used our technique in several different applications, including isolating antibodies against impure proteins and complex antigens, where several rounds of phage display often fail. Our results indicate that antibody arrays can be used to identify differentially expressed proteins.</description><subject>Amino Acid Sequence</subject><subject>Antibodies</subject><subject>Antibodies - chemistry</subject><subject>antibody arrays</subject><subject>Antigen-Antibody Reactions</subject><subject>Antigens</subject><subject>Arrays</subject><subject>Bacteria - chemistry</subject><subject>Bacteria - genetics</subject><subject>Biochemistry - methods</subject><subject>Biological and medical sciences</subject><subject>Biosensing Techniques - methods</subject><subject>Biotechnology</subject><subject>Blotting, Western</subject><subject>Cloning</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Enzymes</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>HeLa Cells</subject><subject>high-throughput screening</subject><subject>Humans</subject><subject>Ligands</subject><subject>Methods. Procedures. Technologies</subject><subject>Molecular Probe Techniques</subject><subject>Molecular Sequence Data</subject><subject>Oligonucleotide Array Sequence Analysis</subject><subject>Others</subject><subject>Peptide Library</subject><subject>Protein Conformation</subject><subject>Proteins</subject><subject>Proteins - chemistry</subject><subject>Proteins - metabolism</subject><subject>Recombinant Proteins - chemistry</subject><subject>Robotics</subject><subject>Serum Albumin - chemistry</subject><subject>Serum Albumin, Bovine - chemistry</subject><subject>Various methods and equipments</subject><issn>1087-0156</issn><issn>1546-1696</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNqN0luL1DAUAOAiintx_4JUcRUfuubS5vI4LF4WFhdc8TWk6UknSycZkxScf2_GDur6oJKHhOTLIefkVNUZRhcYUfGGy1a2D6pj3LWswUyyh2WNBG8Q7thRdZLSHUKItYw9ro4wkpwSQo6rjyufXR-GXa1j1LtU2xDrtRvXTV7HMI_r7ZzrZCKAd36sg6314UKzX4zga-czRG2yCz49qR5ZPSU4O8yn1e27t58vPzTXN--vLlfXjWFE5IYCFmZAGnqrSTsMAwHWdy2SQg6tGDqDtEDMMCEAgaREaABiO9xZZoHS0-rVEnUbw9cZUlYblwxMk_YQ5qQ4I4QjLHCRL_8uCaGSi-6fEHPWCcpJgc__gHdhjr4kq0pBKeaEtwVdLGjUEyjnbcilQmUMsHEmeLCu7K-wLF8ipdxHfX3vQjEZvuVRzympq9tP_29vvty354s1MaQUwaptdBsddwojte8b9aNvint6yGruNzD8ppZGKeDFAehk9GSj9salX65DbUt4Yc8W5nWeI_w8931G5VGqfDD9DunS0eY</recordid><startdate>20000901</startdate><enddate>20000901</enddate><creator>de Wildt, Ruud M. 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Psychology</topic><topic>HeLa Cells</topic><topic>high-throughput screening</topic><topic>Humans</topic><topic>Ligands</topic><topic>Methods. Procedures. Technologies</topic><topic>Molecular Probe Techniques</topic><topic>Molecular Sequence Data</topic><topic>Oligonucleotide Array Sequence Analysis</topic><topic>Others</topic><topic>Peptide Library</topic><topic>Protein Conformation</topic><topic>Proteins</topic><topic>Proteins - chemistry</topic><topic>Proteins - metabolism</topic><topic>Recombinant Proteins - chemistry</topic><topic>Robotics</topic><topic>Serum Albumin - chemistry</topic><topic>Serum Albumin, Bovine - chemistry</topic><topic>Various methods and equipments</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>de Wildt, Ruud M. 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T</au><au>Tomlinson, Ian M</au><au>Mundy, Chris R</au><au>Gorick, Barbara D</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Antibody arrays for high-throughput screening of antibody-antigen interactions</atitle><jtitle>Nature biotechnology</jtitle><addtitle>Nat Biotechnol</addtitle><date>2000-09-01</date><risdate>2000</risdate><volume>18</volume><issue>9</issue><spage>989</spage><epage>994</epage><pages>989-994</pages><issn>1087-0156</issn><eissn>1546-1696</eissn><coden>NABIF9</coden><abstract>We have developed a novel technique for high-throughput screening of recombinant antibodies, based on the creation of antibody arrays. Our method uses robotic picking and high-density gridding of bacteria containing antibody genes followed by filter-based enzyme-linked immunosorbent assay (ELISA) screening to identify clones that express binding antibody fragments. By eliminating the need for liquid handling, we can thereby screen up to 18,342 different antibody clones at a time and, because the clones are arrayed from master stocks, the same antibodies can be double spotted and screened simultaneously against 15 different antigens. We have used our technique in several different applications, including isolating antibodies against impure proteins and complex antigens, where several rounds of phage display often fail. Our results indicate that antibody arrays can be used to identify differentially expressed proteins.</abstract><cop>New York, NY</cop><pub>Nature</pub><pmid>10973222</pmid><doi>10.1038/79494</doi><tpages>6</tpages></addata></record> |
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| ispartof | Nature biotechnology, 2000-09, Vol.18 (9), p.989-994 |
| issn | 1087-0156 1546-1696 |
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| subjects | Amino Acid Sequence Antibodies Antibodies - chemistry antibody arrays Antigen-Antibody Reactions Antigens Arrays Bacteria - chemistry Bacteria - genetics Biochemistry - methods Biological and medical sciences Biosensing Techniques - methods Biotechnology Blotting, Western Cloning Enzyme-Linked Immunosorbent Assay Enzymes Fundamental and applied biological sciences. Psychology HeLa Cells high-throughput screening Humans Ligands Methods. Procedures. Technologies Molecular Probe Techniques Molecular Sequence Data Oligonucleotide Array Sequence Analysis Others Peptide Library Protein Conformation Proteins Proteins - chemistry Proteins - metabolism Recombinant Proteins - chemistry Robotics Serum Albumin - chemistry Serum Albumin, Bovine - chemistry Various methods and equipments |
| title | Antibody arrays for high-throughput screening of antibody-antigen interactions |
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