title: An improved nonchromatographic method for the purification of recombinant proteins using elastin-like polypeptide-tagged proteases.
abstract: Proteins fused to the elastin-like polypeptide (ELP) tag can be selectively separated from crude cell extract without chromatography. To avoid the interference of the ELP tag on properties of the target protein, it is necessary to remove the ELP tag from target protein by protease digestion. Therefore, an additional chromatographic purification step is required to remove the proteases, and this is time- and labor-consuming. Here we demonstrate the utility of the ELP-tagged proteases for cleavage of ELP fusion proteins, allowing one-step removal of the cleaved ELP tag and ELP-tagged proteases without chromatography.
authors: State Key Laboratory of Biocontrol, Guangdong Province Key Laboratory of Therapeutic Functional Genes, College of Life Sciences, Sun Yat-sen University, Guangdong 510275, People's Republic of China.
authors: Dongming
authors: D
authors: Lan
authors: Guangrui
authors: G
authors: Huang
authors: Hongwei
authors: H
authors: Shao
authors: Lichun
authors: L
authors: Zhang
authors: Lixin
authors: L
authors: Ma
authors: Shangwu
authors: S
authors: Chen
authors: Anlong
authors: A
authors: Xu
references: Protein Eng Des Sel. 2004 Jan;17(1):57-66
references: 14985538
references: Anal Biochem. 2007 Jan 1;360(1):166-8
references: 17084802
references: Prog Biophys Mol Biol. 1992;57(1):23-57
references: 1549698
references: Electrophoresis. 2004 May;25(9):1327-33
references: 15174055
references: Mol Cell. 2000 May;5(5):865-76
references: 10882122
references: Nat Methods. 2005 Sep;2(9):659-61
references: 16074986
references: J Am Chem Soc. 2005 Aug 17;127(32):11228-9
references: 16089436
references: Biotechnol Prog. 2006 May-Jun;22(3):638-46
references: 16739944
references: Nat Biotechnol. 1999 Nov;17(11):1112-5
references: 10545920
references: Biochem J. 2006 Sep 15;398(3):577-83
references: 16716147
references: Biotechnol Bioeng. 2003 Jan 5;81(1):74-9
references: 12432583
references: J Struct Funct Genomics. 2004;5(1-2):75-86
references: 15263846
references: Biomacromolecules. 2006 Sep;7(9):2475-8
references: 16961306
references: Protein Sci. 2004 Dec;13(12):3274-84
references: 15557268
references: J Biol Chem. 1990 Jun 5;265(16):9062-5
references: 2160953
references: Biotechnol Prog. 2001 Jul-Aug;17(4):720-8
references: 11485434
references: J Microbiol Biotechnol. 2007 Nov;17(11):1751-7
references: 18092457