{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,24]],"date-time":"2025-10-24T16:19:36Z","timestamp":1761322776482,"version":"build-2065373602"},"reference-count":36,"publisher":"Wiley","issue":"1-2","license":[{"start":{"date-parts":[[2013,11,27]],"date-time":"2013-11-27T00:00:00Z","timestamp":1385510400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J of Separation Science"],"published-print":{"date-parts":[[2014,1]]},"abstract":"<jats:p>The development of new catechol\u2010<jats:italic><jats:styled-content style=\"fixed-case\">O<\/jats:styled-content><\/jats:italic>\u2010methyltransferase inhibitors has led to an improvement in the treatment of <jats:styled-content style=\"fixed-case\">P<\/jats:styled-content>arkinson's disease. However, despite the fact that the soluble isoform has been extensively investigated, few studies have been published concerning membrane isoform chromatographic recovery and bioactivity levels. In this work, chromatographic profiles of both catechol\u2010<jats:italic><jats:styled-content style=\"fixed-case\">O<\/jats:styled-content><\/jats:italic>\u2010methyltransferase isoforms were compared using quaternary amine as a ligand to evaluate its activity levels and recovery rates. Results show that both proteins required different conditions for adsorption; the soluble isoform adsorption was performed at low ionic strength, while the membrane isoform required increasing linear salt gradient. However, the application of 0.5% <jats:styled-content style=\"fixed-case\">T<\/jats:styled-content>riton <jats:styled-content style=\"fixed-case\">X<\/jats:styled-content>\u2010100 promoted membrane isoform adsorption even at low ionic strength. Indeed, chromatographic conditions of both isoforms became similar when detergents were applied. The developed methods also appear to be highly effective in bioactivity recovery, presenting rates of 107% for soluble protein and 67 and 91% for membrane isoform without and with detergents, respectively. The chromatographic strategies with and without detergents resulted in a 4.3\u2010 and sevenfold purification, respectively, corresponding to specific activity values of 331 and 496\u00a0nmol\/h\/mg. Thus, the use of <jats:styled-content style=\"fixed-case\">Q<\/jats:styled-content>\u2010sepharose as anion exchanger was effective in the recovery of both enzymes, which is a requirement for further kinetic and pharmacological trials.<\/jats:p>","DOI":"10.1002\/jssc.201300977","type":"journal-article","created":{"date-parts":[[2013,10,31]],"date-time":"2013-10-31T06:27:26Z","timestamp":1383200846000},"page":"20-29","source":"Crossref","is-referenced-by-count":9,"title":["Recovery of biological active catechol\u2010<i><scp>O<\/scp><\/i>\u2010methyltransferase isoforms from <scp>Q<\/scp>\u2010sepharose"],"prefix":"10.1002","volume":"37","author":[{"given":"F. F.","family":"Correia","sequence":"first","affiliation":[{"name":"CICS\u2010UBI\u2010Centro de Investiga\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade Universidade da Beira Interior  Covilh\u00e3 Portugal"}]},{"given":"F. M.","family":"Santos","sequence":"additional","affiliation":[{"name":"CICS\u2010UBI\u2010Centro de Investiga\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade Universidade da Beira Interior  Covilh\u00e3 Portugal"}]},{"given":"A. Q.","family":"Pedro","sequence":"additional","affiliation":[{"name":"CICS\u2010UBI\u2010Centro de Investiga\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade Universidade da Beira Interior  Covilh\u00e3 Portugal"}]},{"given":"M. J.","family":"Bonif\u00e1cio","sequence":"additional","affiliation":[{"name":"Departamento de Investigac\u00e3o e Desenvolvimento BIAL  S. Mamede do Coronado Portugal"}]},{"given":"J. A.","family":"Queiroz","sequence":"additional","affiliation":[{"name":"CICS\u2010UBI\u2010Centro de Investiga\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade Universidade da Beira Interior  Covilh\u00e3 Portugal"}]},{"given":"L. A.","family":"Passarinha","sequence":"additional","affiliation":[{"name":"CICS\u2010UBI\u2010Centro de Investiga\u00e7\u00e3o em Ci\u00eancias da Sa\u00fade Universidade da Beira Interior  Covilh\u00e3 Portugal"}]}],"member":"311","published-online":{"date-parts":[[2013,11,27]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1527-3458.2007.00020.x"},{"key":"e_1_2_7_3_1","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1016\/S0031-6997(24)01423-6","volume":"51","author":"M\u00e4nnist\u00f6 P. T.","year":"1999","journal-title":"Pharmacol. Rev."},{"key":"e_1_2_7_4_1","first-page":"231","volume":"228","author":"Bonif\u00e1cio M. J.","year":"2003","journal-title":"Meth. Mol. Biol."},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-8993(99)01063-X"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2011.08.022"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0163-7258(98)00032-1"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1007\/s40262-012-0024-7"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/bmc.779"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/0378-4347(96)00117-X"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chroma.2007.06.002"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/bmc.1377"},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj1450135"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/0167-4781(92)90479-J"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/0006-291X(91)91517-G"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jchromb.2011.03.056"},{"key":"e_1_2_7_17_1","doi-asserted-by":"publisher","DOI":"10.1385\/1-59259-400-X:195"},{"volume-title":"Membrane Protein Purification and Crystallization","year":"2003","author":"Hunte C.","key":"e_1_2_7_18_1"},{"key":"e_1_2_7_19_1","doi-asserted-by":"publisher","DOI":"10.1080\/10826068.2012.654570"},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.1002\/jssc.201200101"},{"key":"e_1_2_7_21_1","doi-asserted-by":"publisher","DOI":"10.1007\/BFb0036121"},{"key":"e_1_2_7_22_1","doi-asserted-by":"publisher","DOI":"10.1002\/jssc.201300010"},{"key":"e_1_2_7_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2012.03.025"},{"key":"e_1_2_7_24_1","doi-asserted-by":"publisher","DOI":"10.1038\/227680a0"},{"key":"e_1_2_7_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-1656(01)00237-1"},{"key":"e_1_2_7_26_1","doi-asserted-by":"publisher","DOI":"10.1002\/jssc.201000347"},{"key":"e_1_2_7_27_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M401086200"},{"key":"e_1_2_7_28_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja039335h"},{"key":"e_1_2_7_29_1","doi-asserted-by":"publisher","DOI":"10.1021\/bc990152n"},{"key":"e_1_2_7_30_1","doi-asserted-by":"publisher","DOI":"10.1006\/abbi.1998.1005"},{"key":"e_1_2_7_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jlumin.2010.05.008"},{"key":"e_1_2_7_32_1","doi-asserted-by":"publisher","DOI":"10.1002\/jssc.200800503"},{"key":"e_1_2_7_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jchromb.2010.05.028"},{"key":"e_1_2_7_34_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1570-0232(03)00087-4"},{"key":"e_1_2_7_35_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.sbi.2008.07.001"},{"key":"e_1_2_7_36_1","doi-asserted-by":"publisher","DOI":"10.3168\/jds.S0022-0302(94)77060-0"},{"key":"e_1_2_7_37_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0308-8146(03)00203-6"}],"container-title":["Journal of Separation Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fjssc.201300977","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jssc.201300977","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,19]],"date-time":"2025-10-19T12:42:11Z","timestamp":1760877731000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/jssc.201300977"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,11,27]]},"references-count":36,"journal-issue":{"issue":"1-2","published-print":{"date-parts":[[2014,1]]}},"alternative-id":["10.1002\/jssc.201300977"],"URL":"https:\/\/doi.org\/10.1002\/jssc.201300977","archive":["Portico"],"relation":{},"ISSN":["1615-9306","1615-9314"],"issn-type":[{"type":"print","value":"1615-9306"},{"type":"electronic","value":"1615-9314"}],"subject":[],"published":{"date-parts":[[2013,11,27]]}}}