doi: 10.1016/s0378-4347(01)00261-4.
Immobilised metal-ion affinity chromatography purification of histidine-tagged recombinant proteins: a wash step with a low concentration of EDTA
Affiliations
- PMID: 11522055
- DOI: 10.1016/s0378-4347(01)00261-4
Item in Clipboard
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.
.
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.
Similar articles
-
Purification of His-Tagged Proteins.Methods Enzymol. 2015;559:1-15. doi: 10.1016/bs.mie.2014.11.003. Epub 2015 May 4. Methods Enzymol. 2015. PMID: 26096499
-
Immobilized metal-ion affinity chromatography of recombinant Fab protein OPG C11 in the presence of EDTA-Mg(II).J Chromatogr A. 2002 Nov 29;978(1-2):153-64. doi: 10.1016/s0021-9673(02)01429-2. J Chromatogr A. 2002. PMID: 12458952
-
Immobilized metal affinity chromatography optimization for poly-histidine tagged proteins.J Chromatogr A. 2020 Oct 11;1629:461505. doi: 10.1016/j.chroma.2020.461505. Epub 2020 Aug 21. J Chromatogr A. 2020. PMID: 32861092
-
Perspectives of immobilized-metal affinity chromatography.J Biochem Biophys Methods. 2001 Oct 30;49(1-3):335-60. doi: 10.1016/s0165-022x(01)00207-x. J Biochem Biophys Methods. 2001. PMID: 11694288 Review.
-
Exploiting the interactions between poly-histidine fusion tags and immobilized metal ions.Biotechnol Lett. 2011 Jun;33(6):1075-84. doi: 10.1007/s10529-011-0554-3. Epub 2011 Feb 12. Biotechnol Lett. 2011. PMID: 21318632 Review.
Cited by
-
Profiling and quantitative evaluation of three nickel-coated magnetic matrices for purification of recombinant proteins: helpful hints for the optimized nanomagnetisable matrix preparation.J Nanobiotechnology. 2011 Aug 8;9:31. doi: 10.1186/1477-3155-9-31. J Nanobiotechnology. 2011. PMID: 21824404 Free PMC article.
-
Simple and Efficient Purification of Recombinant Proteins Using the Heparin-Binding Affinity Tag.Curr Protoc Protein Sci. 2017 Nov 1;90:6.16.1-6.16.13. doi: 10.1002/cpps.41. Curr Protoc Protein Sci. 2017. PMID: 29091276 Free PMC article.
-
Fully automated protein purification.Anal Biochem. 2009 Oct 15;393(2):176-81. doi: 10.1016/j.ab.2009.07.009. Epub 2009 Jul 28. Anal Biochem. 2009. PMID: 19595984 Free PMC article.
-
One Step Purification-Vaccine Delivery System.Pharmaceutics. 2023 May 1;15(5):1390. doi: 10.3390/pharmaceutics15051390. Pharmaceutics. 2023. PMID: 37242632 Free PMC article.
-
Challenges and opportunities in the purification of recombinant tagged proteins.Biotechnol Adv. 2014 Mar-Apr;32(2):366-81. doi: 10.1016/j.biotechadv.2013.12.001. Epub 2013 Dec 12. Biotechnol Adv. 2014. PMID: 24334194 Free PMC article. Review.
MeSH terms
Substances
LinkOut - more resources
Other Literature Sources