Aptamer-Based Alternatives to the Conventional Immobilized Metal Affinity Chromatography for Purification of His-Tagged Proteins

Autor:	Hyun Kyung Lim, Hye Yeon Nam, Sang Soo Hah, Il-Hyun Kim, Seonmi Shin
Rok vydání:	2013
Předmět:	Tandem affinity purification Strep-tag Chromatography Oligonucleotide Chemistry Aptamer Biochemistry (medical) Clinical Biochemistry Ion chromatography Biochemistry Analytical Chemistry Molecular recognition Affinity chromatography Electrochemistry Affinity electrophoresis Spectroscopy
Zdroj:	Analytical Letters. 46:407-415
ISSN:	1532-236X 0003-2719
DOI:	10.1080/00032719.2012.721105
Popis:	Aptamers are oligonucleotides or peptide molecules that are able to bind to their specific target molecules with high affinity via molecular recognition. In this study, we present development of aptamer-based affinity purification for His-tagged proteins for comparison of purification efficiency with the conventional Ni2+-based affinity chromatography. Thiol-functionalized aptamers able to specifically bind to His-tag were immobilized employing two crosslinking methods onto the surface of polystyrene resins. The resulting aptamer-anchored resins were successfully applied for purification of His-tagged proteins from complex E. coli and human cell lysates, respectively, and superior or at least comparable purification results to the conventional immobilized metal affinity chromatography were obtained via one-step purification.
Databáze:	OpenAIRE
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