NDLI: A Benzothiazolium Salt as Chromatography Ligand for Protein Purification

Content Provider	Springer Nature Link
Author	Alves, L. P. Ramos, S. S. Sousa, F. Boto, R. E. F. Almeida, P.
Copyright Year	2014
Abstract	A new chromatography support was synthesized from Sepharose CL-6B containing a benzothiazolium bromide salt as a ligand. The original goal of this study was the evaluation of the azolium salt individual contribution as a constituent part of thiacarbocyanine dyes in an affinity or multi-modal relationship with standard proteins. This dye was previously post-grafted onto beaded cellulose by a curing method and successfully used as an affinity ligand for the interaction and isolation of different proteins. Following this idea, the immobilization of an azolium bromide salt onto Sepharose CL-6B in different concentrations was performed by Steglich esterification, using dicyclohexylcarbodiimide as a coupling reagent and 4-dimethylaminopyridine as basic catalyzer. The obtained crafted beaded Sepharose supports were qualitatively and quantitatively characterized by scanning electron microscopy, energy dispersive X-ray spectroscopy and elemental analysis. The interactions between the chromatographic supports obtained and several standard proteins were finally evaluated. The best results were achieved using a chromatographic support with a ligand density of 0.18 mmol of benzothiazolium salt/g of chromatography support, enabling the separation of ribonuclease, α-chymotrypsin and bovine serum albumin from an artificial mixture using a decreasing ammonium sulfate stepwise gradient. In conclusion, this study demonstrates the potential applicability of small multi-modal cationic ligands such as benzothiazolium salts, compared to these congener cyanine dyes previously studied, for the separation and purification of proteins by chromatography, revealing a distinct and useful chromatographic behavior by itself.
Starting Page	1597
Ending Page	1605
Page Count	9
File Format	PDF
ISSN	00095893
Journal	Chromatographia
Volume Number	77
Issue Number	23-24
e-ISSN	16121112
Language	English
Publisher	Springer Berlin Heidelberg
Publisher Date	2014-10-15
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Multi-modal chromatography Proteins purification Steglich esterification Benzothiazolium salt Chromatography Proteomics Pharmacy Laboratory Medicine
Content Type	Text
Resource Type	Article
Subject	Organic Chemistry Analytical Chemistry Biochemistry Clinical Biochemistry

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