title: Rhodamine B as ligand for affinity chromatography: chromatographic studies on derivatized beaded cellulose.
abstract: Rhodamine B (RB) post-grafted onto beaded cellulose by a curing method is used as a biomimetic ligand in dye affinity chromatography. The grafted materials obtained are qualitatively characterized by scanning electron microscope and fourier transformed infrared spectroscopy. An amount of 76.79 mmol RB/g dyed cellulose is determined by elemental analysis. The RB affinity interaction with the trypsin, alpha-chymotrypsin, and BSA (bovine serum albumin) is analyzed using different mobile phase composition. The results show a selective separation of a mixture of BSA and trypsin into two single peaks by step elution with 1.75 M, 0.5 M, and 0 M ammonium sulphate in the eluent buffer. A good reproducibility of the retention time is obtained for these proteins in the mixture with typical values of 8.0 +/- 0.2 min for BSA and 20.0 +/- 0.2 min for trypsin, showing a possible application in the purification of samples with different composition.
authors.affiliation: Departamento de Química and Unidade de I & D de Materiais Têxteis e Papeleiros, Universidade da Beira Interior, PT-6201-001 Covilhã, Portugal.
authors.forename: Andreia
authors.initials: A
authors.lastname: Silva
authors.forename: Renato E F
authors.initials: RE
authors.lastname: Boto
authors.forename: Cândida T
authors.initials: CT
authors.lastname: Tomaz
authors.forename: João A
authors.initials: JA
authors.lastname: Queiroz
authors.forename: Paulo
authors.initials: P
authors.lastname: Almeida
mesh_terms: D000818:Animals; D002417:Cattle; D002482:Cellulose / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D002918:Chymotrypsin / Q000378:metabolism; D008024:Ligands; D011485:Protein Binding; D012235:Rhodamines / Q000737:chemistry* / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
chemical_list: D008024:Ligands; D012235:Rhodamines; D012710:Serum Albumin, Bovine; D002482:Cellulose; D002918:Chymotrypsin; C117608:alpha-chymotrypsin; D014357:Trypsin; C029773:rhodamine B