{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,19]],"date-time":"2025-10-19T15:45:57Z","timestamp":1760888757123,"version":"build-2065373602"},"reference-count":23,"publisher":"Wiley","issue":"22","license":[{"start":{"date-parts":[[2012,11,23]],"date-time":"2012-11-23T00:00:00Z","timestamp":1353628800000},"content-version":"vor","delay-in-days":22,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"Portuguese Foundation for Science and Technology","award":["PTDC\/EQU-EQU\/"],"award-info":[{"award-number":["PTDC\/EQU-EQU\/"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J of Separation Science"],"published-print":{"date-parts":[[2012,11]]},"abstract":"<jats:p>This study addresses the feasibility of scaling\u2010up the removal of host cell impurities from plasmid <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> (p<jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content>)\u2010containing <jats:italic><jats:styled-content style=\"fixed-case\">E<\/jats:styled-content>scherichia coli<\/jats:italic> lysates by phenyl\u2010boronic (<jats:styled-content style=\"fixed-case\">PB<\/jats:styled-content>) acid chromatography using columns packed with 7.6 and 15.2 cm<jats:sup>3<\/jats:sup> of controlled porous glass beads (<jats:styled-content style=\"fixed-case\">CPG<\/jats:styled-content>) derivatized with <jats:styled-content style=\"fixed-case\">PB<\/jats:styled-content> ligands. Equilibration was performed with water at 10 cm<jats:sup>3<\/jats:sup>\/min and no conditioning of the lysate feed was required. At a ratio of lysate feed to adsorbent volume of 1.3, 93\u201396% of p<jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> was recovered in the flow through while 66\u201371% of impurities remained bound (\u223c2.5\u2010fold purification). The entire sequence of loading, washing, elution, and re\u2010equilibration was completed in 20 min. Run\u2010to\u2010run consistency was observed in terms of chromatogram features and performance (yield, purification factor, agarose electrophoresis) across the different amounts of adsorbent (0.75\u201315.2 cm<jats:sup>3<\/jats:sup>) by performing successive injections of lysates prepared independently and containing 3.7 or 6.1 kbp plasmids. The column productivity at large scale was 4 dm<jats:sup>3<\/jats:sup> of alkaline lysate per hour per dm<jats:sup>3<\/jats:sup> of <jats:styled-content style=\"fixed-case\">PB<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">CPG<\/jats:styled-content> resin. The method is rapid, reproducible, simple, and straightforward to scale\u2010up. Furthermore, it is capable of handling heavily contaminated samples, constituting a good alternative to purification techniques such as isopropanol precipitation, aqueous two\u2010phase systems, and tangential flow filtration.<\/jats:p>","DOI":"10.1002\/jssc.201200225","type":"journal-article","created":{"date-parts":[[2012,11,23]],"date-time":"2012-11-23T02:29:48Z","timestamp":1353637788000},"page":"3190-3196","source":"Crossref","is-referenced-by-count":6,"title":["Validation and scale\u2010up of plasmid <scp>DNA<\/scp> purification by phenyl\u2010boronic acid chromatography"],"prefix":"10.1002","volume":"35","author":[{"given":"A. Gabriela","family":"Gomes","sequence":"first","affiliation":[{"name":"IBB Institute for Biotechnology and Bioengineering Centre for Biological and Chemical Engineering Department of Bioengineering Instituto Superior T\u00e9cnico Universidade T\u00e9cnica de Lisboa Lisboa Portugal"}]},{"given":"Ana M.","family":"Azevedo","sequence":"additional","affiliation":[{"name":"IBB Institute for Biotechnology and Bioengineering Centre for Biological and Chemical Engineering Department of Bioengineering Instituto Superior T\u00e9cnico Universidade T\u00e9cnica de Lisboa Lisboa Portugal"}]},{"given":"M. Raquel","family":"Aires\u2010Barros","sequence":"additional","affiliation":[{"name":"IBB Institute for Biotechnology and Bioengineering Centre for Biological and Chemical Engineering Department of Bioengineering Instituto Superior T\u00e9cnico Universidade T\u00e9cnica de Lisboa Lisboa Portugal"}]},{"given":"D. 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