Rhodamine B as ligand for affinity chromatography: chromatographic studies on derivatized beaded cellulose.
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.
Silva
Andreia
A
Departamento de Química and Unidade de I & D de Materiais Têxteis e Papeleiros, Universidade da Beira Interior, PT-6201-001 Covilhã, Portugal.
Boto
Renato E F
RE
Tomaz
Cândida T
CT
Queiroz
João A
JA
Almeida
Paulo
P
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
D008024:Ligands; D012235:Rhodamines; D012710:Serum Albumin, Bovine; D002482:Cellulose; D002918:Chymotrypsin; C117608:alpha-chymotrypsin; D014357:Trypsin; C029773:rhodamine B