Immobilized anthraquinone dyes for affinity chromatography

✍ Scribed by Robert S. Beissner; Frederick B. Rudolph

• Publisher: Elsevier Science
• Year: 1978
• Tongue: English
• Weight: 724 KB
• Volume: 161
• Category: Article
• ISSN: 1873-3778
• DOI: 10.1016/s0021-9673(01)85220-1

✦ Synopsis

Cibacron Brilliant Blue BR-P, a structural isomer of Cibacron Blue 3G-A (the chromophore in Blue Dextran), and Procion Brilliant Blue MR, Cibacron-Brilliant. Blue BR-P without the sulfonated bet& moiety, were covalently attached to Sepharose CL-6B by the triazine rings. The use of these materials for column af&ity chromatography *was compared with commercial Blue Sepharose CL-6B, Cibacron Blue 3G-A covalently attached by its triazine ring to Sepharose CL-6B, and with a column of agarose succinyl-polyacrylic hydrazine with Cibacron Blue 3G-A attached on its anthraquinone ring. It was found likely that the I-amino-anthraquinone-2sulfonic acid moiety was the critical structure for binding of the selected enzymes used, one kinase and three dehydrogenases. These enzymes, bound to the dyes, , attached to the columns by their triazine rings and elution of the enzymes occurred at similar concentrations of eluting buffers indicating that the entire Cibacron Blue 3G-A molecule is not required for biospecific binding and elution. The new columns described in this report may-allow for selective enzyme pur&ation due to differences in binding and steric interactions of the proteins with the different dyes.