{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,23]],"date-time":"2025-10-23T16:46:19Z","timestamp":1761237979299,"version":"build-2065373602"},"reference-count":32,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2013,5,3]],"date-time":"2013-05-03T00:00:00Z","timestamp":1367539200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"University of Beira Interior-Health Sciences Research Centre"},{"name":"FCT","award":["EXPL\/BBB-BQB\/0960\/2012","SFRH\/BD\/81222\/2011","Pest-C\/SAU\/UI0709\/2011"],"award-info":[{"award-number":["EXPL\/BBB-BQB\/0960\/2012","SFRH\/BD\/81222\/2011","Pest-C\/SAU\/UI0709\/2011"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J of Separation Science"],"published-print":{"date-parts":[[2013,6]]},"abstract":"Despite of membrane catechol\u2010O<\/jats:italic>\u2010methyltransferase (MBCOMT<\/jats:styled-content>, EC<\/jats:styled-content> 2.1.1.6) physiological importance on catecholamines\u2019 O<\/jats:styled-content><\/jats:italic>\u2010methylation, no studies allowed their total isolation. Therefore, for the first time, we compare the performance of three hydrophobic adsorbents (butyl\u2010, epoxy\u2010, and octyl\u2010Sepharose) in purification of recombinant human COMT<\/jats:styled-content> (hMBCOMT<\/jats:styled-content>) from crude B<\/jats:styled-content>revibacillus choshinensis<\/jats:italic> cell lysates to develop a sustainable chromatographic process. Hydrophobic matrices were evaluated in terms of selectivity and hMBCOMT<\/jats:styled-content>'s binding and elution conditions. Results show that hMBCOMT<\/jats:styled-content>'s adsorption was promoted on octyl and butyl at \u2264375 mM NaH2<\/jats:sub>PO4,<\/jats:sub> while on epoxy higher concentrations (>850 mM) were required. Additionally, hMBCOMT<\/jats:styled-content>'s elution was promoted on epoxy, butyl, and octyl using respectively 0.1\u20130.5, 0.25\u20131, and 1% of T<\/jats:styled-content>riton X<\/jats:styled-content>\u2010100. On butyl media, a stepwise strategy using 375 and 0 mM N<\/jats:styled-content>aH<\/jats:styled-content>2<\/jats:sub>PO<\/jats:styled-content>4<\/jats:sub>, followed by three elution steps at 0.25, 0.7 and 1% T<\/jats:styled-content>riton X<\/jats:styled-content>\u2010100, allowed selective hMBCOMT<\/jats:styled-content> isolation. In conclusion, significant amounts of MB<\/jats:styled-content>COMT<\/jats:styled-content> were purified with high selectivity on a single chromatography procedure, despite its elution occurs on multiple peaks. Although successful applications of hydrophobic interaction chromatography in purification of membrane proteins are uncommon, we proved that traditional hydrophobic matrices can open a promising unexplored field to fulfill specific requirements for kinetic and pharmacological trials.<\/jats:p>","DOI":"10.1002\/jssc.201300010","type":"journal-article","created":{"date-parts":[[2013,3,14]],"date-time":"2013-03-14T04:03:27Z","timestamp":1363233807000},"page":"1693-1702","source":"Crossref","is-referenced-by-count":10,"title":["Performance of hydrophobic interaction ligands for human membrane\u2010bound catechol\u2010O<\/i>\u2010methyltransferase purification"],"prefix":"10.1002","volume":"36","author":[{"given":"F\u00e1tima Milhano","family":"Santos","sequence":"first","affiliation":[{"name":"CICS\u2010UBI\u2010 Centro de Investiga\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade Universidade da Beira Interior Covilh\u00e3 Portugal"}]},{"given":"Augusto Quaresma","family":"Pedro","sequence":"additional","affiliation":[{"name":"CICS\u2010UBI\u2010 Centro de Investiga\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade Universidade da Beira Interior Covilh\u00e3 Portugal"}]},{"given":"Rui Filipe","family":"Soares","sequence":"additional","affiliation":[{"name":"CICS\u2010UBI\u2010 Centro de Investiga\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade Universidade da Beira Interior Covilh\u00e3 Portugal"}]},{"given":"Rita","family":"Martins","sequence":"additional","affiliation":[{"name":"CICS\u2010UBI\u2010 Centro de Investiga\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade Universidade da Beira Interior Covilh\u00e3 Portugal"}]},{"given":"Maria Jo\u00e3o","family":"Bonif\u00e1cio","sequence":"additional","affiliation":[{"name":"Departamento de Investiga\u00e7\u00e3o e Desenvolvimento BIAL S. 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