Oligonucleotide-assisted cleavage and ligation: a novel directional DNA cloning technology to capture cDNAs. Application in the construction of a human immune antibody phage-display library
Identifieur interne : 000F05 ( Istex/Corpus ); précédent : 000F04; suivant : 000F06Oligonucleotide-assisted cleavage and ligation: a novel directional DNA cloning technology to capture cDNAs. Application in the construction of a human immune antibody phage-display library
Auteurs : Sonia Schoonbroodt ; Nicolas Frans ; Mark Desouza ; Rachel Eren ; Smadar Priel ; Naama Brosh ; Judith Ben-Porath ; Arie Zauberman ; Ehud Ilan ; Shlomo Dagan ; Edward H. Cohen ; Hennie R. Hoogenboom ; Robert Charles Ladner ; Rene M. HoetSource :
- Nucleic Acids Research [ 0305-1048 ] ; 2005.
Abstract
The use of oligonucleotide-assisted cleavage and ligation (ONCL), a novel approach to the capture of gene repertoires, in the construction of a phage-display immune antibody library is described. ONCL begins with rapid amplification of cDNA ends to amplify all members equally. A single, specific cut near 5′ and/or 3′ end of each gene fragment (in single stranded form) is facilitated by hybridization with an appropriate oligonucleotide adapter. Directional cloning of targeted DNA is accomplished by ligation of a partially duplex DNA molecule (containing suitable restriction sites) and amplification with primers in constant regions. To demonstrate utility and reliability of ONCL, a human antibody repertoire was cloned from IgG mRNA extracted from human B-lymphocytes engrafted in Trimera mice. These mice were transplanted with peripheral blood lymphocytes from Candida albicans infected individuals and subsequently immunized with C.albicans glyceraldehyde-3-phosphate dehydrogenase (GAPDH). DNA sequencing showed that ONCL resulted in efficient capture of gene repertoires. Indeed, full representation of all VH families/segments was observed showing that ONCL did not introduce cloning biases for or against any VH family. We validated the efficiency of ONCL by creating a functional Fab phage-display library with a size of 3.3 × 1010 and by selecting five unique Fabs against GAPDH antigen.
Url:
DOI: 10.1093/nar/gni080
Links to Exploration step
ISTEX:9A6B8EE691D0FDF2B5ACBD67FAD5EF122CF10A32Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Oligonucleotide-assisted cleavage and ligation: a novel directional DNA cloning technology to capture cDNAs. Application in the construction of a human immune antibody phage-display library</title><author><name sortKey="Schoonbroodt, Sonia" sort="Schoonbroodt, Sonia" uniqKey="Schoonbroodt S" first="Sonia" last="Schoonbroodt">Sonia Schoonbroodt</name></author><author><name sortKey="Frans, Nicolas" sort="Frans, Nicolas" uniqKey="Frans N" first="Nicolas" last="Frans">Nicolas Frans</name></author><author><name sortKey="Desouza, Mark" sort="Desouza, Mark" uniqKey="Desouza M" first="Mark" last="Desouza">Mark Desouza</name><affiliation><mods:affiliation>Dyax Corp., Corporate Headquarters 300 Technology Square, Cambridge, MA 02139, USA</mods:affiliation></affiliation></author><author><name sortKey="Eren, Rachel" sort="Eren, Rachel" uniqKey="Eren R" first="Rachel" last="Eren">Rachel Eren</name><affiliation><mods:affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</mods:affiliation></affiliation></author><author><name sortKey="Priel, Smadar" sort="Priel, Smadar" uniqKey="Priel S" first="Smadar" last="Priel">Smadar Priel</name><affiliation><mods:affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</mods:affiliation></affiliation></author><author><name sortKey="Brosh, Naama" sort="Brosh, Naama" uniqKey="Brosh N" first="Naama" last="Brosh">Naama Brosh</name><affiliation><mods:affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</mods:affiliation></affiliation></author><author><name sortKey="Ben Porath, Judith" sort="Ben Porath, Judith" uniqKey="Ben Porath J" first="Judith" last="Ben-Porath">Judith Ben-Porath</name><affiliation><mods:affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</mods:affiliation></affiliation></author><author><name sortKey="Zauberman, Arie" sort="Zauberman, Arie" uniqKey="Zauberman A" first="Arie" last="Zauberman">Arie Zauberman</name><affiliation><mods:affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</mods:affiliation></affiliation></author><author><name sortKey="Ilan, Ehud" sort="Ilan, Ehud" uniqKey="Ilan E" first="Ehud" last="Ilan">Ehud Ilan</name><affiliation><mods:affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</mods:affiliation></affiliation></author><author><name sortKey="Dagan, Shlomo" sort="Dagan, Shlomo" uniqKey="Dagan S" first="Shlomo" last="Dagan">Shlomo Dagan</name><affiliation><mods:affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</mods:affiliation></affiliation></author><author><name sortKey="Cohen, Edward H" sort="Cohen, Edward H" uniqKey="Cohen E" first="Edward H." last="Cohen">Edward H. Cohen</name><affiliation><mods:affiliation>Dyax Corp., Corporate Headquarters 300 Technology Square, Cambridge, MA 02139, USA</mods:affiliation></affiliation></author><author><name sortKey="Hoogenboom, Hennie R" sort="Hoogenboom, Hennie R" uniqKey="Hoogenboom H" first="Hennie R." last="Hoogenboom">Hennie R. Hoogenboom</name></author><author><name sortKey="Ladner, Robert Charles" sort="Ladner, Robert Charles" uniqKey="Ladner R" first="Robert Charles" last="Ladner">Robert Charles Ladner</name><affiliation><mods:affiliation>Dyax Corp., Corporate Headquarters 300 Technology Square, Cambridge, MA 02139, USA</mods:affiliation></affiliation></author><author><name sortKey="Hoet, Rene M" sort="Hoet, Rene M" uniqKey="Hoet R" first="Rene M." last="Hoet">Rene M. Hoet</name></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:9A6B8EE691D0FDF2B5ACBD67FAD5EF122CF10A32</idno><date when="2005" year="2005">2005</date><idno type="doi">10.1093/nar/gni080</idno><idno type="url">https://api.istex.fr/document/9A6B8EE691D0FDF2B5ACBD67FAD5EF122CF10A32/fulltext/pdf</idno><idno type="wicri:Area/Istex/Corpus">000F05</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Oligonucleotide-assisted cleavage and ligation: a novel directional DNA cloning technology to capture cDNAs. Application in the construction of a human immune antibody phage-display library</title><author><name sortKey="Schoonbroodt, Sonia" sort="Schoonbroodt, Sonia" uniqKey="Schoonbroodt S" first="Sonia" last="Schoonbroodt">Sonia Schoonbroodt</name></author><author><name sortKey="Frans, Nicolas" sort="Frans, Nicolas" uniqKey="Frans N" first="Nicolas" last="Frans">Nicolas Frans</name></author><author><name sortKey="Desouza, Mark" sort="Desouza, Mark" uniqKey="Desouza M" first="Mark" last="Desouza">Mark Desouza</name><affiliation><mods:affiliation>Dyax Corp., Corporate Headquarters 300 Technology Square, Cambridge, MA 02139, USA</mods:affiliation></affiliation></author><author><name sortKey="Eren, Rachel" sort="Eren, Rachel" uniqKey="Eren R" first="Rachel" last="Eren">Rachel Eren</name><affiliation><mods:affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</mods:affiliation></affiliation></author><author><name sortKey="Priel, Smadar" sort="Priel, Smadar" uniqKey="Priel S" first="Smadar" last="Priel">Smadar Priel</name><affiliation><mods:affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</mods:affiliation></affiliation></author><author><name sortKey="Brosh, Naama" sort="Brosh, Naama" uniqKey="Brosh N" first="Naama" last="Brosh">Naama Brosh</name><affiliation><mods:affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</mods:affiliation></affiliation></author><author><name sortKey="Ben Porath, Judith" sort="Ben Porath, Judith" uniqKey="Ben Porath J" first="Judith" last="Ben-Porath">Judith Ben-Porath</name><affiliation><mods:affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</mods:affiliation></affiliation></author><author><name sortKey="Zauberman, Arie" sort="Zauberman, Arie" uniqKey="Zauberman A" first="Arie" last="Zauberman">Arie Zauberman</name><affiliation><mods:affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</mods:affiliation></affiliation></author><author><name sortKey="Ilan, Ehud" sort="Ilan, Ehud" uniqKey="Ilan E" first="Ehud" last="Ilan">Ehud Ilan</name><affiliation><mods:affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</mods:affiliation></affiliation></author><author><name sortKey="Dagan, Shlomo" sort="Dagan, Shlomo" uniqKey="Dagan S" first="Shlomo" last="Dagan">Shlomo Dagan</name><affiliation><mods:affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</mods:affiliation></affiliation></author><author><name sortKey="Cohen, Edward H" sort="Cohen, Edward H" uniqKey="Cohen E" first="Edward H." last="Cohen">Edward H. Cohen</name><affiliation><mods:affiliation>Dyax Corp., Corporate Headquarters 300 Technology Square, Cambridge, MA 02139, USA</mods:affiliation></affiliation></author><author><name sortKey="Hoogenboom, Hennie R" sort="Hoogenboom, Hennie R" uniqKey="Hoogenboom H" first="Hennie R." last="Hoogenboom">Hennie R. Hoogenboom</name></author><author><name sortKey="Ladner, Robert Charles" sort="Ladner, Robert Charles" uniqKey="Ladner R" first="Robert Charles" last="Ladner">Robert Charles Ladner</name><affiliation><mods:affiliation>Dyax Corp., Corporate Headquarters 300 Technology Square, Cambridge, MA 02139, USA</mods:affiliation></affiliation></author><author><name sortKey="Hoet, Rene M" sort="Hoet, Rene M" uniqKey="Hoet R" first="Rene M." last="Hoet">Rene M. Hoet</name></author></analytic><monogr></monogr><series><title level="j">Nucleic Acids Research</title><title level="j" type="abbrev">Nucl. Acids Res.</title><idno type="ISSN">0305-1048</idno><idno type="eISSN">1362-4962</idno><imprint><publisher>Oxford University Press</publisher><date type="published" when="2005">2005</date><biblScope unit="volume">33</biblScope><biblScope unit="issue">9</biblScope><biblScope unit="page" from="81">e81</biblScope><biblScope unit="page" to="81">e81</biblScope></imprint><idno type="ISSN">0305-1048</idno></series><idno type="istex">9A6B8EE691D0FDF2B5ACBD67FAD5EF122CF10A32</idno><idno type="DOI">10.1093/nar/gni080</idno><idno type="local">gni080</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0305-1048</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The use of oligonucleotide-assisted cleavage and ligation (ONCL), a novel approach to the capture of gene repertoires, in the construction of a phage-display immune antibody library is described. ONCL begins with rapid amplification of cDNA ends to amplify all members equally. A single, specific cut near 5′ and/or 3′ end of each gene fragment (in single stranded form) is facilitated by hybridization with an appropriate oligonucleotide adapter. Directional cloning of targeted DNA is accomplished by ligation of a partially duplex DNA molecule (containing suitable restriction sites) and amplification with primers in constant regions. To demonstrate utility and reliability of ONCL, a human antibody repertoire was cloned from IgG mRNA extracted from human B-lymphocytes engrafted in Trimera mice. These mice were transplanted with peripheral blood lymphocytes from Candida albicans infected individuals and subsequently immunized with C.albicans glyceraldehyde-3-phosphate dehydrogenase (GAPDH). DNA sequencing showed that ONCL resulted in efficient capture of gene repertoires. Indeed, full representation of all VH families/segments was observed showing that ONCL did not introduce cloning biases for or against any VH family. We validated the efficiency of ONCL by creating a functional Fab phage-display library with a size of 3.3 × 1010 and by selecting five unique Fabs against GAPDH antigen.</div></front></TEI><istex><corpusName>oup</corpusName><author><json:item><name>Sonia Schoonbroodt</name></json:item><json:item><name>Nicolas Frans</name></json:item><json:item><name>Mark DeSouza</name><affiliations><json:string>Dyax Corp., Corporate Headquarters 300 Technology Square, Cambridge, MA 02139, USA</json:string></affiliations></json:item><json:item><name>Rachel Eren</name><affiliations><json:string>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</json:string></affiliations></json:item><json:item><name>Smadar Priel</name><affiliations><json:string>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</json:string></affiliations></json:item><json:item><name>Naama Brosh</name><affiliations><json:string>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</json:string></affiliations></json:item><json:item><name>Judith Ben-Porath</name><affiliations><json:string>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</json:string></affiliations></json:item><json:item><name>Arie Zauberman</name><affiliations><json:string>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</json:string></affiliations></json:item><json:item><name>Ehud Ilan</name><affiliations><json:string>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</json:string></affiliations></json:item><json:item><name>Shlomo Dagan</name><affiliations><json:string>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</json:string></affiliations></json:item><json:item><name>Edward H. Cohen</name><affiliations><json:string>Dyax Corp., Corporate Headquarters 300 Technology Square, Cambridge, MA 02139, USA</json:string></affiliations></json:item><json:item><name>Hennie R. Hoogenboom</name></json:item><json:item><name>Robert Charles Ladner</name><affiliations><json:string>Dyax Corp., Corporate Headquarters 300 Technology Square, Cambridge, MA 02139, USA</json:string></affiliations></json:item><json:item><name>René M. Hoet</name><affiliations><json:string>E-mail: rhoet@dyax.com</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>1</value></json:item></subject><language><json:string>eng</json:string></language><originalGenre><json:string>other</json:string></originalGenre><abstract>The use of oligonucleotide-assisted cleavage and ligation (ONCL), a novel approach to the capture of gene repertoires, in the construction of a phage-display immune antibody library is described. ONCL begins with rapid amplification of cDNA ends to amplify all members equally. A single, specific cut near 5′ and/or 3′ end of each gene fragment (in single stranded form) is facilitated by hybridization with an appropriate oligonucleotide adapter. Directional cloning of targeted DNA is accomplished by ligation of a partially duplex DNA molecule (containing suitable restriction sites) and amplification with primers in constant regions. To demonstrate utility and reliability of ONCL, a human antibody repertoire was cloned from IgG mRNA extracted from human B-lymphocytes engrafted in Trimera mice. These mice were transplanted with peripheral blood lymphocytes from Candida albicans infected individuals and subsequently immunized with C.albicans glyceraldehyde-3-phosphate dehydrogenase (GAPDH). DNA sequencing showed that ONCL resulted in efficient capture of gene repertoires. Indeed, full representation of all VH families/segments was observed showing that ONCL did not introduce cloning biases for or against any VH family. We validated the efficiency of ONCL by creating a functional Fab phage-display library with a size of 3.3 × 1010 and by selecting five unique Fabs against GAPDH antigen.</abstract><qualityIndicators><score>7.924</score><pdfVersion>1.4</pdfVersion><pdfPageSize>612 x 794 pts</pdfPageSize><refBibsNative>false</refBibsNative><keywordCount>1</keywordCount><abstractCharCount>1403</abstractCharCount><pdfWordCount>9070</pdfWordCount><pdfCharCount>57940</pdfCharCount><pdfPageCount>14</pdfPageCount><abstractWordCount>202</abstractWordCount></qualityIndicators><title>Oligonucleotide-assisted cleavage and ligation: a novel directional DNA cloning technology to capture cDNAs. Application in the construction of a human immune antibody phage-display library</title><genre><json:string>other</json:string></genre><host><volume>33</volume><publisherId><json:string>nar</json:string></publisherId><pages><last>e81</last><first>e81</first></pages><issn><json:string>0305-1048</json:string></issn><issue>9</issue><genre><json:string>journal</json:string></genre><language><json:string>unknown</json:string></language><eissn><json:string>1362-4962</json:string></eissn><title>Nucleic Acids Research</title></host><categories><wos><json:string>BIOCHEMISTRY & MOLECULAR BIOLOGY</json:string></wos></categories><publicationDate>2005</publicationDate><copyrightDate>2005</copyrightDate><doi><json:string>10.1093/nar/gni080</json:string></doi><id>9A6B8EE691D0FDF2B5ACBD67FAD5EF122CF10A32</id><score>0.24773598</score><fulltext><json:item><original>true</original><mimetype>application/pdf</mimetype><extension>pdf</extension><uri>https://api.istex.fr/document/9A6B8EE691D0FDF2B5ACBD67FAD5EF122CF10A32/fulltext/pdf</uri></json:item><json:item><original>false</original><mimetype>application/zip</mimetype><extension>zip</extension><uri>https://api.istex.fr/document/9A6B8EE691D0FDF2B5ACBD67FAD5EF122CF10A32/fulltext/zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/document/9A6B8EE691D0FDF2B5ACBD67FAD5EF122CF10A32/fulltext/tei"><teiHeader><fileDesc><titleStmt><title level="a" type="main" xml:lang="en">Oligonucleotide-assisted cleavage and ligation: a novel directional DNA cloning technology to capture cDNAs. Application in the construction of a human immune antibody phage-display library</title><respStmt><resp>Références bibliographiques récupérées via GROBID</resp><name resp="ISTEX-API">ISTEX-API (INIST-CNRS)</name></respStmt></titleStmt><publicationStmt><authority>ISTEX</authority><publisher>Oxford University Press</publisher><availability><p>© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oupjournals.org</p></availability><date>2005</date></publicationStmt><notesStmt><note>*To whom correspondence should be addressed. Tel: +32 4 364 2405; Fax: +32 4 364 2499; Email: rhoet@dyax.com</note></notesStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a" type="main" xml:lang="en">Oligonucleotide-assisted cleavage and ligation: a novel directional DNA cloning technology to capture cDNAs. Application in the construction of a human immune antibody phage-display library</title><author xml:id="author-1"><persName><forename type="first">Sonia</forename><surname>Schoonbroodt</surname></persName></author><author xml:id="author-2"><persName><forename type="first">Nicolas</forename><surname>Frans</surname></persName></author><author xml:id="author-3"><persName><forename type="first">Mark</forename><surname>DeSouza</surname></persName><affiliation>Dyax Corp., Corporate Headquarters 300 Technology Square, Cambridge, MA 02139, USA</affiliation></author><author xml:id="author-4"><persName><forename type="first">Rachel</forename><surname>Eren</surname></persName><affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</affiliation></author><author xml:id="author-5"><persName><forename type="first">Smadar</forename><surname>Priel</surname></persName><affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</affiliation></author><author xml:id="author-6"><persName><forename type="first">Naama</forename><surname>Brosh</surname></persName><affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</affiliation></author><author xml:id="author-7"><persName><forename type="first">Judith</forename><surname>Ben-Porath</surname></persName><affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</affiliation></author><author xml:id="author-8"><persName><forename type="first">Arie</forename><surname>Zauberman</surname></persName><affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</affiliation></author><author xml:id="author-9"><persName><forename type="first">Ehud</forename><surname>Ilan</surname></persName><affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</affiliation></author><author xml:id="author-10"><persName><forename type="first">Shlomo</forename><surname>Dagan</surname></persName><affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</affiliation></author><author xml:id="author-11"><persName><forename type="first">Edward H.</forename><surname>Cohen</surname></persName><affiliation>Dyax Corp., Corporate Headquarters 300 Technology Square, Cambridge, MA 02139, USA</affiliation></author><author xml:id="author-12"><persName><forename type="first">Hennie R.</forename><surname>Hoogenboom</surname></persName></author><author xml:id="author-13"><persName><forename type="first">Robert Charles</forename><surname>Ladner</surname></persName><affiliation>Dyax Corp., Corporate Headquarters 300 Technology Square, Cambridge, MA 02139, USA</affiliation></author><author xml:id="author-14"><persName><forename type="first">René M.</forename><surname>Hoet</surname></persName></author></analytic><monogr><title level="j">Nucleic Acids Research</title><title level="j" type="abbrev">Nucl. Acids Res.</title><idno type="pISSN">0305-1048</idno><idno type="eISSN">1362-4962</idno><imprint><publisher>Oxford University Press</publisher><date type="published" when="2005"></date><biblScope unit="volume">33</biblScope><biblScope unit="issue">9</biblScope><biblScope unit="page" from="81">e81</biblScope><biblScope unit="page" to="81">e81</biblScope></imprint></monogr><idno type="istex">9A6B8EE691D0FDF2B5ACBD67FAD5EF122CF10A32</idno><idno type="DOI">10.1093/nar/gni080</idno><idno type="local">gni080</idno></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>2005</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>The use of oligonucleotide-assisted cleavage and ligation (ONCL), a novel approach to the capture of gene repertoires, in the construction of a phage-display immune antibody library is described. ONCL begins with rapid amplification of cDNA ends to amplify all members equally. A single, specific cut near 5′ and/or 3′ end of each gene fragment (in single stranded form) is facilitated by hybridization with an appropriate oligonucleotide adapter. Directional cloning of targeted DNA is accomplished by ligation of a partially duplex DNA molecule (containing suitable restriction sites) and amplification with primers in constant regions. To demonstrate utility and reliability of ONCL, a human antibody repertoire was cloned from IgG mRNA extracted from human B-lymphocytes engrafted in Trimera mice. These mice were transplanted with peripheral blood lymphocytes from Candida albicans infected individuals and subsequently immunized with C.albicans glyceraldehyde-3-phosphate dehydrogenase (GAPDH). DNA sequencing showed that ONCL resulted in efficient capture of gene repertoires. Indeed, full representation of all VH families/segments was observed showing that ONCL did not introduce cloning biases for or against any VH family. We validated the efficiency of ONCL by creating a functional Fab phage-display library with a size of 3.3 × 1010 and by selecting five unique Fabs against GAPDH antigen.</p></abstract><textClass><keywords scheme="keyword"><list><head>hwp-journal-coll</head><item><term>1</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="2005">Published</change><change xml:id="refBibs-istex" who="#ISTEX-API" when="2016-10-14">References added</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><original>false</original><mimetype>text/plain</mimetype><extension>txt</extension><uri>https://api.istex.fr/document/9A6B8EE691D0FDF2B5ACBD67FAD5EF122CF10A32/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="corpus oup" wicri:toSee="no header"><istex:xmlDeclaration>version="1.0" encoding="US-ASCII"</istex:xmlDeclaration><istex:docType PUBLIC="-//NLM//DTD Journal Publishing DTD v2.3 20070202//EN" URI="journalpublishing.dtd" name="istex:docType"></istex:docType><istex:document><article xml:lang="en" article-type="other"><front><journal-meta><journal-id journal-id-type="hwp">nar</journal-id><journal-id journal-id-type="nlm-ta">Nucleic Acids Res</journal-id><journal-id journal-id-type="publisher-id">nar</journal-id><journal-title>Nucleic Acids Research</journal-title><abbrev-journal-title abbrev-type="publisher">Nucl. Acids Res.</abbrev-journal-title><issn pub-type="ppub">0305-1048</issn><issn pub-type="epub">1362-4962</issn><publisher><publisher-name>Oxford University Press</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="other">gni080</article-id><article-id pub-id-type="doi">10.1093/nar/gni080</article-id><article-categories><subj-group subj-group-type="heading"><subject>Methods Online</subject></subj-group><subj-group subj-group-type="hwp-journal-coll"><subject>1</subject></subj-group></article-categories><title-group><article-title>Oligonucleotide-assisted cleavage and ligation: a novel directional DNA cloning technology to capture cDNAs. Application in the construction of a human immune antibody phage-display library</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Schoonbroodt</surname><given-names>Sonia</given-names></name></contrib><contrib contrib-type="author"><name><surname>Frans</surname><given-names>Nicolas</given-names></name></contrib><contrib contrib-type="author"><name><surname>DeSouza</surname><given-names>Mark</given-names></name><xref rid="AU1">1</xref></contrib><contrib contrib-type="author"><name><surname>Eren</surname><given-names>Rachel</given-names></name><xref rid="AU2">2</xref></contrib><contrib contrib-type="author"><name><surname>Priel</surname><given-names>Smadar</given-names></name><xref rid="AU2">2</xref></contrib><contrib contrib-type="author"><name><surname>Brosh</surname><given-names>Naama</given-names></name><xref rid="AU2">2</xref></contrib><contrib contrib-type="author"><name><surname>Ben-Porath</surname><given-names>Judith</given-names></name><xref rid="AU2">2</xref></contrib><contrib contrib-type="author"><name><surname>Zauberman</surname><given-names>Arie</given-names></name><xref rid="AU2">2</xref></contrib><contrib contrib-type="author"><name><surname>Ilan</surname><given-names>Ehud</given-names></name><xref rid="AU2">2</xref></contrib><contrib contrib-type="author"><name><surname>Dagan</surname><given-names>Shlomo</given-names></name><xref rid="AU2">2</xref></contrib><contrib contrib-type="author"><name><surname>Cohen</surname><given-names>Edward H.</given-names></name><xref rid="AU1">1</xref></contrib><contrib contrib-type="author"><name><surname>Hoogenboom</surname><given-names>Hennie R.</given-names></name></contrib><contrib contrib-type="author"><name><surname>Ladner</surname><given-names>Robert Charles</given-names></name><xref rid="AU1">1</xref></contrib><contrib contrib-type="author"><name><surname>Hoet</surname><given-names>René M.</given-names></name><xref rid="COR1">*</xref></contrib><aff>Dyax s.a., Boulevard du Rectorat 27B Sart Tilman, B-4000 Liege 1, Belgium</aff><aff id="AU1"><sup>1</sup>Dyax Corp., Corporate Headquarters 300 Technology Square, Cambridge, MA 02139, USA</aff><aff id="AU2"><sup>2</sup>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</aff></contrib-group><author-notes><corresp id="COR1"><sup>*</sup>To whom correspondence should be addressed. Tel: +32 4 364 2405; Fax: +32 4 364 2499; Email: <ext-link xlink:href="rhoet@dyax.com" ext-link-type="email">rhoet@dyax.com</ext-link></corresp></author-notes><pub-date pub-type="ppub"><year>2005</year></pub-date><volume>33</volume><issue>9</issue><fpage>e81</fpage><lpage>e81</lpage><history><date date-type="accepted"><day>28</day><month>4</month><year>2005</year></date><date date-type="received"><day>7</day><month>12</month><year>2004</year></date><date date-type="rev-recd"><day>29</day><month>3</month><year>2005</year></date></history><permissions><copyright-statement>© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact <ext-link xlink:href="journals.permissions@oupjournals.org" ext-link-type="email">journals.permissions@oupjournals.org</ext-link></copyright-statement><copyright-year>2005</copyright-year><license license-type="open-access"><p></p></license></permissions><abstract xml:lang="en"><p>The use of oligonucleotide-assisted cleavage and ligation (ONCL), a novel approach to the capture of gene repertoires, in the construction of a phage-display immune antibody library is described. ONCL begins with rapid amplification of cDNA ends to amplify all members equally. A single, specific cut near 5′ and/or 3′ end of each gene fragment (in single stranded form) is facilitated by hybridization with an appropriate oligonucleotide adapter. Directional cloning of targeted DNA is accomplished by ligation of a partially duplex DNA molecule (containing suitable restriction sites) and amplification with primers in constant regions. To demonstrate utility and reliability of ONCL, a human antibody repertoire was cloned from IgG mRNA extracted from human B-lymphocytes engrafted in Trimera mice. These mice were transplanted with peripheral blood lymphocytes from <italic>Candida albicans</italic> infected individuals and subsequently immunized with <italic>C.albicans</italic> glyceraldehyde-3-phosphate dehydrogenase (GAPDH). DNA sequencing showed that ONCL resulted in efficient capture of gene repertoires. Indeed, full representation of all V<sub>H</sub> families/segments was observed showing that ONCL did not introduce cloning biases for or against any V<sub>H</sub> family. We validated the efficiency of ONCL by creating a functional Fab phage-display library with a size of 3.3 × 10<sup>10</sup> and by selecting five unique Fabs against GAPDH antigen.</p></abstract><custom-meta-wrap><custom-meta><meta-name>hwp-legacy-fpage</meta-name><meta-value>e81</meta-value></custom-meta><custom-meta><meta-name>cover-date</meta-name><meta-value></meta-value></custom-meta><custom-meta><meta-name>hwp-legacy-dochead</meta-name><meta-value>research-article</meta-value></custom-meta></custom-meta-wrap></article-meta></front></article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Oligonucleotide-assisted cleavage and ligation: a novel directional DNA cloning technology to capture cDNAs. Application in the construction of a human immune antibody phage-display library</title></titleInfo><titleInfo type="alternative" lang="en" contentType="CDATA"><title>Oligonucleotide-assisted cleavage and ligation: a novel directional DNA cloning technology to capture cDNAs. Application in the construction of a human immune antibody phage-display library</title></titleInfo><name type="personal"><namePart type="given">Sonia</namePart><namePart type="family">Schoonbroodt</namePart><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Nicolas</namePart><namePart type="family">Frans</namePart><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Mark</namePart><namePart type="family">DeSouza</namePart><affiliation>Dyax Corp., Corporate Headquarters 300 Technology Square, Cambridge, MA 02139, USA</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Rachel</namePart><namePart type="family">Eren</namePart><affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Smadar</namePart><namePart type="family">Priel</namePart><affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Naama</namePart><namePart type="family">Brosh</namePart><affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Judith</namePart><namePart type="family">Ben-Porath</namePart><affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Arie</namePart><namePart type="family">Zauberman</namePart><affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Ehud</namePart><namePart type="family">Ilan</namePart><affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Shlomo</namePart><namePart type="family">Dagan</namePart><affiliation>XTL Biopharmaceuticals Ltd Kiryat Weizmann, Rehovot, 76103, Israel</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Edward H.</namePart><namePart type="family">Cohen</namePart><affiliation>Dyax Corp., Corporate Headquarters 300 Technology Square, Cambridge, MA 02139, USA</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Hennie R.</namePart><namePart type="family">Hoogenboom</namePart><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Robert Charles</namePart><namePart type="family">Ladner</namePart><affiliation>Dyax Corp., Corporate Headquarters 300 Technology Square, Cambridge, MA 02139, USA</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">René M.</namePart><namePart type="family">Hoet</namePart><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="other" displayLabel="other"></genre><subject><genre>hwp-journal-coll</genre><topic>1</topic></subject><originInfo><publisher>Oxford University Press</publisher><dateIssued encoding="w3cdtf">2005</dateIssued><copyrightDate encoding="w3cdtf">2005</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType></physicalDescription><abstract lang="en">The use of oligonucleotide-assisted cleavage and ligation (ONCL), a novel approach to the capture of gene repertoires, in the construction of a phage-display immune antibody library is described. ONCL begins with rapid amplification of cDNA ends to amplify all members equally. A single, specific cut near 5′ and/or 3′ end of each gene fragment (in single stranded form) is facilitated by hybridization with an appropriate oligonucleotide adapter. Directional cloning of targeted DNA is accomplished by ligation of a partially duplex DNA molecule (containing suitable restriction sites) and amplification with primers in constant regions. To demonstrate utility and reliability of ONCL, a human antibody repertoire was cloned from IgG mRNA extracted from human B-lymphocytes engrafted in Trimera mice. These mice were transplanted with peripheral blood lymphocytes from Candida albicans infected individuals and subsequently immunized with C.albicans glyceraldehyde-3-phosphate dehydrogenase (GAPDH). DNA sequencing showed that ONCL resulted in efficient capture of gene repertoires. Indeed, full representation of all VH families/segments was observed showing that ONCL did not introduce cloning biases for or against any VH family. We validated the efficiency of ONCL by creating a functional Fab phage-display library with a size of 3.3 × 1010 and by selecting five unique Fabs against GAPDH antigen.</abstract><note type="author-notes">*To whom correspondence should be addressed. Tel: +32 4 364 2405; Fax: +32 4 364 2499; Email: rhoet@dyax.com</note><relatedItem type="host"><titleInfo><title>Nucleic Acids Research</title></titleInfo><titleInfo type="abbreviated"><title>Nucl. Acids Res.</title></titleInfo><genre type="journal">journal</genre><identifier type="ISSN">0305-1048</identifier><identifier type="eISSN">1362-4962</identifier><identifier type="PublisherID">nar</identifier><identifier type="PublisherID-hwp">nar</identifier><identifier type="PublisherID-nlm-ta">Nucleic Acids Res</identifier><part><date>2005</date><detail type="volume"><caption>vol.</caption><number>33</number></detail><detail type="issue"><caption>no.</caption><number>9</number></detail><extent unit="pages"><start>e81</start><end>e81</end></extent></part></relatedItem><identifier type="istex">9A6B8EE691D0FDF2B5ACBD67FAD5EF122CF10A32</identifier><identifier type="DOI">10.1093/nar/gni080</identifier><identifier type="local">gni080</identifier><accessCondition type="use and reproduction" contentType="copyright">© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oupjournals.org</accessCondition><recordInfo><recordContentSource>OUP</recordContentSource></recordInfo></mods></metadata><annexes><json:item><original>true</original><mimetype>image/jpeg</mimetype><extension>jpeg</extension><uri>https://api.istex.fr/document/9A6B8EE691D0FDF2B5ACBD67FAD5EF122CF10A32/annexes/jpeg</uri></json:item><json:item><original>true</original><mimetype>image/gif</mimetype><extension>gif</extension><uri>https://api.istex.fr/document/9A6B8EE691D0FDF2B5ACBD67FAD5EF122CF10A32/annexes/gif</uri></json:item></annexes><enrichments><istex:catWosTEI uri="https://api.istex.fr/document/9A6B8EE691D0FDF2B5ACBD67FAD5EF122CF10A32/enrichments/catWos"><teiHeader><profileDesc><textClass><classCode scheme="WOS">BIOCHEMISTRY & MOLECULAR BIOLOGY</classCode></textClass></profileDesc></teiHeader></istex:catWosTEI><json:item><type>refBibs</type><uri>https://api.istex.fr/document/9A6B8EE691D0FDF2B5ACBD67FAD5EF122CF10A32/enrichments/refBibs</uri></json:item></enrichments><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Wicri/Belgique/explor/OpenAccessBelV2/Data/Istex/Corpus
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000F05 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Istex/Corpus/biblio.hfd -nk 000F05 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Wicri/Belgique
|area= OpenAccessBelV2
|flux= Istex
|étape= Corpus
|type= RBID
|clé= ISTEX:9A6B8EE691D0FDF2B5ACBD67FAD5EF122CF10A32
|texte= Oligonucleotide-assisted cleavage and ligation: a novel directional DNA cloning technology to capture cDNAs. Application in the construction of a human immune antibody phage-display library
}}
| This area was generated with Dilib version V0.6.25. |  |