Details
| Original language | English |
|---|---|
| Pages (from-to) | 203-213 |
| Number of pages | 11 |
| Journal | Journal of Chromatography A |
| Volume | 855 |
| Issue number | 1 |
| Publication status | Published - 3 Sept 1999 |
| Externally published | Yes |
Abstract
A procedure for renaturation of heterodimeric platelet-derived growth factor (PDGF-AB) from inclusion bodies of recombinant Escherichia coli using size-exclusion chromatography is described. Either prepurified or crude PDGF-AB inclusion bodies solubilized with guanidinium hydrochloride were subjected to buffer exchange from denaturing to renaturing conditions during chromatography. Renaturation of PDGF-AB involves folding of the solubilized and unfolded molecules into dimerization competent monomers during size-exclusion chromatography and subsequent dimerization of folded monomers into the biologically active heterodimeric growth factor. Optimized conditions result in an overall yield of 75% active PDGF-AB with respect to size-exclusion chromatography and subsequent dimerization. The described approach allows renaturation at high protein concentrations and circumvents aggregation which is observed when refolding is carried out by dilution. Copyright (C) 1999 Elsevier Science B.V.
Keywords
- Escherichia coli, Growth factors, Proteins
ASJC Scopus subject areas
- Chemistry(all)
- Analytical Chemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Chemistry(all)
- Organic Chemistry
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In: Journal of Chromatography A, Vol. 855, No. 1, 03.09.1999, p. 203-213.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Renaturation of heterodimeric platelet-derived growth factor from inclusion bodies of recombinant Escherichia coli using size-exclusion chromatography
AU - Müller, Carsten
AU - Rinas, Ursula
PY - 1999/9/3
Y1 - 1999/9/3
N2 - A procedure for renaturation of heterodimeric platelet-derived growth factor (PDGF-AB) from inclusion bodies of recombinant Escherichia coli using size-exclusion chromatography is described. Either prepurified or crude PDGF-AB inclusion bodies solubilized with guanidinium hydrochloride were subjected to buffer exchange from denaturing to renaturing conditions during chromatography. Renaturation of PDGF-AB involves folding of the solubilized and unfolded molecules into dimerization competent monomers during size-exclusion chromatography and subsequent dimerization of folded monomers into the biologically active heterodimeric growth factor. Optimized conditions result in an overall yield of 75% active PDGF-AB with respect to size-exclusion chromatography and subsequent dimerization. The described approach allows renaturation at high protein concentrations and circumvents aggregation which is observed when refolding is carried out by dilution. Copyright (C) 1999 Elsevier Science B.V.
AB - A procedure for renaturation of heterodimeric platelet-derived growth factor (PDGF-AB) from inclusion bodies of recombinant Escherichia coli using size-exclusion chromatography is described. Either prepurified or crude PDGF-AB inclusion bodies solubilized with guanidinium hydrochloride were subjected to buffer exchange from denaturing to renaturing conditions during chromatography. Renaturation of PDGF-AB involves folding of the solubilized and unfolded molecules into dimerization competent monomers during size-exclusion chromatography and subsequent dimerization of folded monomers into the biologically active heterodimeric growth factor. Optimized conditions result in an overall yield of 75% active PDGF-AB with respect to size-exclusion chromatography and subsequent dimerization. The described approach allows renaturation at high protein concentrations and circumvents aggregation which is observed when refolding is carried out by dilution. Copyright (C) 1999 Elsevier Science B.V.
KW - Escherichia coli
KW - Growth factors
KW - Proteins
UR - http://www.scopus.com/inward/record.url?scp=0038227075&partnerID=8YFLogxK
U2 - 10.1016/S0021-9673(99)00660-3
DO - 10.1016/S0021-9673(99)00660-3
M3 - Article
C2 - 10514985
AN - SCOPUS:0038227075
VL - 855
SP - 203
EP - 213
JO - Journal of Chromatography A
JF - Journal of Chromatography A
SN - 0021-9673
IS - 1
ER -