Immobilised metal-ion affinity chromatography purification of histidine-tagged recombinant proteins: a wash step with a low concentration of EDTA.
J Chromatogr B Biomed Sci Appl
; 760(1): 129-36, 2001 Aug 25.
Article
em En
| MEDLINE
| ID: mdl-11522055
ABSTRACT
Immobilised metal-ion affinity chromatography (IMAC) is widely used for the purification of recombinant proteins in which a poly-histidine tag is introduced. However, other proteins may also bind to IMAC columns. We describe the use of a washing buffer with a low concentration of EDTA (0.5 mM) for the removal of proteins without histidine tag from IMAC columns. Four histidine-tagged recombinant proteins/protein complexes were purified to homogeneity from cell culture medium of insect cells by using an EDTA washing buffer. The presence of a low concentration of EDTA in washing buffers during IMAC may have a general application in the purification of histidine-tagged proteins.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas Recombinantes
/
Quelantes
/
Cromatografia de Afinidade
/
Ácido Edético
/
Histidina
Idioma:
En
Revista:
J Chromatogr B Biomed Sci Appl
Assunto da revista:
QUIMICA CLINICA
Ano de publicação:
2001
Tipo de documento:
Article
País de afiliação:
Holanda