Continuous affinity-based selection: rapid screening and simultaneous amplification of bacterial surface-display libraries
A new method for continuous biopanning has been developed. We have combined the power of affinity chromatography with the fecundity of bacteria in a unique process that mimics clonal selection. Mixed populations of bacteria were applied to a fermenter containing the immobilized ligand of interest. B...
Gespeichert in:
| Veröffentlicht in: | Biochemical journal 2001-08, Vol.357 (Pt 3), p.779-785 |
|---|---|
| Hauptverfasser: | , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 785 |
|---|---|
| container_issue | Pt 3 |
| container_start_page | 779 |
| container_title | Biochemical journal |
| container_volume | 357 |
| creator | Patel, D Vitovski, S Senior, H J Edge, M D Hockney, R C Dempsey, M J Sayers, J R |
| description | A new method for continuous biopanning has been developed. We have combined the power of affinity chromatography with the fecundity of bacteria in a unique process that mimics clonal selection. Mixed populations of bacteria were applied to a fermenter containing the immobilized ligand of interest. Bacteria retained in this affinity fermenter were allowed to grow under continuous washout conditions, such that weakly bound organisms were selectively lost. Those initially rare founder bacteria expressing a receptor for the immobilized ligand (R+ve) were thus enriched and amplified simultaneously. From an initial culture containing 1 x 10(10) R-ve cells spiked with fewer than 30 R+ve bacteria ( |
| doi_str_mv | 10.1042/0264-6021:3570779 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_1222007</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>71030403</sourcerecordid><originalsourceid>FETCH-LOGICAL-c457t-84035265c696b67ba657eaa1ca75fda68e20d1389db2227df34220ea06d3772b3</originalsourceid><addsrcrecordid>eNqFkk9v1DAQxa0KRLeFD8AF5cQtMP4TO9sDElq1FKkSFzhbE2fcGjlOsBOk5dOTZVcFTpUPlj2_9-TxG8Zec3jHQYn3ILSqNQh-JRsDxmzP2IYrA3VrRPuMbR7r5-yilO8AXIGCF-ycc6WlVO2G_dqNaQ5pGZdSofchhXlfd1iorwpFcnMY01WVcQrrhctEKaT7CtN6CsMSZ0z0RzpMMfjg8MBXo686dDPlgLEqS_boqO5DmSLuqxi6jDlQecmee4yFXp32S_bt5vrr7ra--_Lp8-7jXe1UY-a6VSAboRunt7rTpkPdGELkDk3je9QtCei5bLd9J4QwvZdKCCAE3UtjRCcv2Yej77R0A_WO0pwx2imHAfPejhjs_5UUHuz9-NPy1Q_ArAZvTwZ5_LFQme0QiqMYj81bw0Gu_yqfBAVXjZJb_iTI23U1ollBfgRdHkvJ5B-fzcEeZsAeMraHjO1pBlbNm3_7_as4hS5_A6e2r6k</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>18181525</pqid></control><display><type>article</type><title>Continuous affinity-based selection: rapid screening and simultaneous amplification of bacterial surface-display libraries</title><source>MEDLINE</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>PubMed Central</source><source>Alma/SFX Local Collection</source><creator>Patel, D ; Vitovski, S ; Senior, H J ; Edge, M D ; Hockney, R C ; Dempsey, M J ; Sayers, J R</creator><creatorcontrib>Patel, D ; Vitovski, S ; Senior, H J ; Edge, M D ; Hockney, R C ; Dempsey, M J ; Sayers, J R</creatorcontrib><description>A new method for continuous biopanning has been developed. We have combined the power of affinity chromatography with the fecundity of bacteria in a unique process that mimics clonal selection. Mixed populations of bacteria were applied to a fermenter containing the immobilized ligand of interest. Bacteria retained in this affinity fermenter were allowed to grow under continuous washout conditions, such that weakly bound organisms were selectively lost. Those initially rare founder bacteria expressing a receptor for the immobilized ligand (R+ve) were thus enriched and amplified simultaneously. From an initial culture containing 1 x 10(10) R-ve cells spiked with fewer than 30 R+ve bacteria (<1 in 10(8)), final ratios of R+ve/R-ve bacteria as high as 1 in 12 were observed, representing an enrichment factor of 55 million-fold. This technology has considerable potential for rapid screening of bacterial surface-display libraries and in facilitating directed-evolution studies.</description><identifier>ISSN: 0264-6021</identifier><identifier>EISSN: 1470-8728</identifier><identifier>DOI: 10.1042/0264-6021:3570779</identifier><identifier>PMID: 11463348</identifier><language>eng</language><publisher>England</publisher><subject>Affinity chromatography ; Bacterial Outer Membrane Proteins - analysis ; Bacterial Proteins ; Chromatography, Affinity - methods ; Escherichia coli - chemistry ; Fermentation ; Fermenters ; Ligands ; Membrane Glycoproteins - analysis ; Peptide Library</subject><ispartof>Biochemical journal, 2001-08, Vol.357 (Pt 3), p.779-785</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c457t-84035265c696b67ba657eaa1ca75fda68e20d1389db2227df34220ea06d3772b3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1222007/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1222007/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,315,728,781,785,886,27929,27930,53796,53798</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11463348$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Patel, D</creatorcontrib><creatorcontrib>Vitovski, S</creatorcontrib><creatorcontrib>Senior, H J</creatorcontrib><creatorcontrib>Edge, M D</creatorcontrib><creatorcontrib>Hockney, R C</creatorcontrib><creatorcontrib>Dempsey, M J</creatorcontrib><creatorcontrib>Sayers, J R</creatorcontrib><title>Continuous affinity-based selection: rapid screening and simultaneous amplification of bacterial surface-display libraries</title><title>Biochemical journal</title><addtitle>Biochem J</addtitle><description>A new method for continuous biopanning has been developed. We have combined the power of affinity chromatography with the fecundity of bacteria in a unique process that mimics clonal selection. Mixed populations of bacteria were applied to a fermenter containing the immobilized ligand of interest. Bacteria retained in this affinity fermenter were allowed to grow under continuous washout conditions, such that weakly bound organisms were selectively lost. Those initially rare founder bacteria expressing a receptor for the immobilized ligand (R+ve) were thus enriched and amplified simultaneously. From an initial culture containing 1 x 10(10) R-ve cells spiked with fewer than 30 R+ve bacteria (<1 in 10(8)), final ratios of R+ve/R-ve bacteria as high as 1 in 12 were observed, representing an enrichment factor of 55 million-fold. This technology has considerable potential for rapid screening of bacterial surface-display libraries and in facilitating directed-evolution studies.</description><subject>Affinity chromatography</subject><subject>Bacterial Outer Membrane Proteins - analysis</subject><subject>Bacterial Proteins</subject><subject>Chromatography, Affinity - methods</subject><subject>Escherichia coli - chemistry</subject><subject>Fermentation</subject><subject>Fermenters</subject><subject>Ligands</subject><subject>Membrane Glycoproteins - analysis</subject><subject>Peptide Library</subject><issn>0264-6021</issn><issn>1470-8728</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkk9v1DAQxa0KRLeFD8AF5cQtMP4TO9sDElq1FKkSFzhbE2fcGjlOsBOk5dOTZVcFTpUPlj2_9-TxG8Zec3jHQYn3ILSqNQh-JRsDxmzP2IYrA3VrRPuMbR7r5-yilO8AXIGCF-ycc6WlVO2G_dqNaQ5pGZdSofchhXlfd1iorwpFcnMY01WVcQrrhctEKaT7CtN6CsMSZ0z0RzpMMfjg8MBXo686dDPlgLEqS_boqO5DmSLuqxi6jDlQecmee4yFXp32S_bt5vrr7ra--_Lp8-7jXe1UY-a6VSAboRunt7rTpkPdGELkDk3je9QtCei5bLd9J4QwvZdKCCAE3UtjRCcv2Yej77R0A_WO0pwx2imHAfPejhjs_5UUHuz9-NPy1Q_ArAZvTwZ5_LFQme0QiqMYj81bw0Gu_yqfBAVXjZJb_iTI23U1ollBfgRdHkvJ5B-fzcEeZsAeMraHjO1pBlbNm3_7_as4hS5_A6e2r6k</recordid><startdate>20010801</startdate><enddate>20010801</enddate><creator>Patel, D</creator><creator>Vitovski, S</creator><creator>Senior, H J</creator><creator>Edge, M D</creator><creator>Hockney, R C</creator><creator>Dempsey, M J</creator><creator>Sayers, J R</creator><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>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20010801</creationdate><title>Continuous affinity-based selection: rapid screening and simultaneous amplification of bacterial surface-display libraries</title><author>Patel, D ; Vitovski, S ; Senior, H J ; Edge, M D ; Hockney, R C ; Dempsey, M J ; Sayers, J R</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c457t-84035265c696b67ba657eaa1ca75fda68e20d1389db2227df34220ea06d3772b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Affinity chromatography</topic><topic>Bacterial Outer Membrane Proteins - analysis</topic><topic>Bacterial Proteins</topic><topic>Chromatography, Affinity - methods</topic><topic>Escherichia coli - chemistry</topic><topic>Fermentation</topic><topic>Fermenters</topic><topic>Ligands</topic><topic>Membrane Glycoproteins - analysis</topic><topic>Peptide Library</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Patel, D</creatorcontrib><creatorcontrib>Vitovski, S</creatorcontrib><creatorcontrib>Senior, H J</creatorcontrib><creatorcontrib>Edge, M D</creatorcontrib><creatorcontrib>Hockney, R C</creatorcontrib><creatorcontrib>Dempsey, M J</creatorcontrib><creatorcontrib>Sayers, J R</creatorcontrib><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>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Biochemical journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Patel, D</au><au>Vitovski, S</au><au>Senior, H J</au><au>Edge, M D</au><au>Hockney, R C</au><au>Dempsey, M J</au><au>Sayers, J R</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Continuous affinity-based selection: rapid screening and simultaneous amplification of bacterial surface-display libraries</atitle><jtitle>Biochemical journal</jtitle><addtitle>Biochem J</addtitle><date>2001-08-01</date><risdate>2001</risdate><volume>357</volume><issue>Pt 3</issue><spage>779</spage><epage>785</epage><pages>779-785</pages><issn>0264-6021</issn><eissn>1470-8728</eissn><abstract>A new method for continuous biopanning has been developed. We have combined the power of affinity chromatography with the fecundity of bacteria in a unique process that mimics clonal selection. Mixed populations of bacteria were applied to a fermenter containing the immobilized ligand of interest. Bacteria retained in this affinity fermenter were allowed to grow under continuous washout conditions, such that weakly bound organisms were selectively lost. Those initially rare founder bacteria expressing a receptor for the immobilized ligand (R+ve) were thus enriched and amplified simultaneously. From an initial culture containing 1 x 10(10) R-ve cells spiked with fewer than 30 R+ve bacteria (<1 in 10(8)), final ratios of R+ve/R-ve bacteria as high as 1 in 12 were observed, representing an enrichment factor of 55 million-fold. This technology has considerable potential for rapid screening of bacterial surface-display libraries and in facilitating directed-evolution studies.</abstract><cop>England</cop><pmid>11463348</pmid><doi>10.1042/0264-6021:3570779</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0264-6021 |
| ispartof | Biochemical journal, 2001-08, Vol.357 (Pt 3), p.779-785 |
| issn | 0264-6021 1470-8728 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_1222007 |
| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Alma/SFX Local Collection |
| subjects | Affinity chromatography Bacterial Outer Membrane Proteins - analysis Bacterial Proteins Chromatography, Affinity - methods Escherichia coli - chemistry Fermentation Fermenters Ligands Membrane Glycoproteins - analysis Peptide Library |
| title | Continuous affinity-based selection: rapid screening and simultaneous amplification of bacterial surface-display libraries |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-12T03%3A24%3A33IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Continuous%20affinity-based%20selection:%20rapid%20screening%20and%20simultaneous%20amplification%20of%20bacterial%20surface-display%20libraries&rft.jtitle=Biochemical%20journal&rft.au=Patel,%20D&rft.date=2001-08-01&rft.volume=357&rft.issue=Pt%203&rft.spage=779&rft.epage=785&rft.pages=779-785&rft.issn=0264-6021&rft.eissn=1470-8728&rft_id=info:doi/10.1042/0264-6021:3570779&rft_dat=%3Cproquest_pubme%3E71030403%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=18181525&rft_id=info:pmid/11463348&rfr_iscdi=true |