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Dye—ligand affinity chromatography on continuous beds
Author(s) -
Mohammad Jamil,
Zeerak Asadullah,
Hjertén Stellan
Publication year - 1995
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1130090205
Subject(s) - chemistry , chromatography , affinity chromatography , triazine , elution , lactate dehydrogenase , alcohol dehydrogenase , dehydrogenase , cofactor , enzyme , biochemistry , organic chemistry
Continuous beds derivatized with three triazine dyes (Cibacron Blue 3G‐A, Pricon Red HE‐3B and Pricon Red H‐3B) were used for chromatographic purification of dehydrogenases from yeast enzyme concentrate. All three columns, which were prepared by a very simple and cost‐effective method, provided strong binding of glucose‐6‐phosphate dehydrogenase and lactate dehydrogenase. The Cibacron Blue 3G‐A column showed high affinity for alcohol dehydrogenase. Under the same conditions, the Pricon Red HE‐3B column showed a lower affinity and the Pricon Red H‐3B column showed none. The adsorbed dehydrogenases were eluted specifically from the columns in high yields (71–113% by desorption with the coenzymes NADP, NADH and NAD respectively). Non‐specific binding of human serum albumin and transferrin to these columns was also investigated. Enzyme assays and analyses by capillary electrophoresis showed that the continuous beds derivatized with triazine dyes gave a high degree of purification.