Renaturation of heterodimeric platelet-derived growth factor from inclusion bodies of recombinant Escherichia coli using size-exclusion chromatography

J Chromatogr A. 1999 Sep 3;855(1):203-13. doi: 10.1016/s0021-9673(99)00660-3.

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.

MeSH terms

  • Chromatography, Gel / methods*
  • Dimerization
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / chemistry*
  • Escherichia coli / genetics
  • Inclusion Bodies / chemistry*
  • Platelet-Derived Growth Factor / chemistry*
  • Protein Renaturation
  • Recombination, Genetic
  • Spectrometry, Fluorescence

Substances

  • Platelet-Derived Growth Factor
  • platelet-derived growth factor AB