Affinity Purification of a Recombinant Protein Expressed as a Fusion with the Maltose-Binding Protein (MBP) Tag

Krisna C. Duong-Ly; Sandra B. Gabelli

doi:10.1016/bs.mie.2014.11.004

Affinity Purification of a Recombinant Protein Expressed as a Fusion with the Maltose-Binding Protein (MBP) Tag

Krisna C. Duong-Ly, Sandra B. Gabelli

School of Medicine

Research output: Chapter in Book/Report/Conference proceeding › Chapter

7 Scopus citations

Abstract

Expression of fusion proteins such as MBP fusions can be used as a way to improve the solubility of the expressed protein in E. coli (Fox and Waugh, 2003; Nallamsetty et al., 2005; Nallamsetty and Waugh, 2006) and as a way to introduce an affinity purification tag. The protocol that follows was designed by the authors as a first step in the purification of a recombinant protein fused with MBP, using fast protein liquid chromatography (FPLC). Cells should have been thawed, resuspended in binding buffer, and lysed by sonication or microfluidization before mixing with the amylose resin or loading on the column. Slight modifications to this protocol may be made to accommodate both the protein of interest and the availability of equipment.

Original language	English (US)
Title of host publication	Methods in Enzymology
Publisher	Academic Press Inc.
Pages	17-26
Number of pages	10
DOIs	https://doi.org/10.1016/bs.mie.2014.11.004
State	Published - Jun 20 2015

Publication series

Name	Methods in Enzymology
Volume	559
ISSN (Print)	0076-6879
ISSN (Electronic)	1557-7988

Keywords

Affinity chromatography
Affinity purification
Fast protein liquid chromatography (FPLC)
Fusion protein
MBP fusion
Maltose-binding protein (MBP)
Recombinant protein

ASJC Scopus subject areas

Biochemistry
Molecular Biology

Access to Document

10.1016/bs.mie.2014.11.004

Cite this

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title = "Affinity Purification of a Recombinant Protein Expressed as a Fusion with the Maltose-Binding Protein (MBP) Tag",

abstract = "Expression of fusion proteins such as MBP fusions can be used as a way to improve the solubility of the expressed protein in E. coli (Fox and Waugh, 2003; Nallamsetty et al., 2005; Nallamsetty and Waugh, 2006) and as a way to introduce an affinity purification tag. The protocol that follows was designed by the authors as a first step in the purification of a recombinant protein fused with MBP, using fast protein liquid chromatography (FPLC). Cells should have been thawed, resuspended in binding buffer, and lysed by sonication or microfluidization before mixing with the amylose resin or loading on the column. Slight modifications to this protocol may be made to accommodate both the protein of interest and the availability of equipment.",

keywords = "Affinity chromatography, Affinity purification, Fast protein liquid chromatography (FPLC), Fusion protein, MBP fusion, Maltose-binding protein (MBP), Recombinant protein",

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AU - Gabelli, Sandra B.

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N2 - Expression of fusion proteins such as MBP fusions can be used as a way to improve the solubility of the expressed protein in E. coli (Fox and Waugh, 2003; Nallamsetty et al., 2005; Nallamsetty and Waugh, 2006) and as a way to introduce an affinity purification tag. The protocol that follows was designed by the authors as a first step in the purification of a recombinant protein fused with MBP, using fast protein liquid chromatography (FPLC). Cells should have been thawed, resuspended in binding buffer, and lysed by sonication or microfluidization before mixing with the amylose resin or loading on the column. Slight modifications to this protocol may be made to accommodate both the protein of interest and the availability of equipment.

AB - Expression of fusion proteins such as MBP fusions can be used as a way to improve the solubility of the expressed protein in E. coli (Fox and Waugh, 2003; Nallamsetty et al., 2005; Nallamsetty and Waugh, 2006) and as a way to introduce an affinity purification tag. The protocol that follows was designed by the authors as a first step in the purification of a recombinant protein fused with MBP, using fast protein liquid chromatography (FPLC). Cells should have been thawed, resuspended in binding buffer, and lysed by sonication or microfluidization before mixing with the amylose resin or loading on the column. Slight modifications to this protocol may be made to accommodate both the protein of interest and the availability of equipment.

KW - Affinity chromatography

KW - Affinity purification

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KW - Fusion protein

KW - MBP fusion

KW - Maltose-binding protein (MBP)

KW - Recombinant protein

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Affinity Purification of a Recombinant Protein Expressed as a Fusion with the Maltose-Binding Protein (MBP) Tag

Abstract

Publication series

Keywords

ASJC Scopus subject areas

Access to Document

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