Recovery of mouse growth hormone from E. coli inclusion bodies using a mild solubilisation and repeated freeze-thaw approach.

ABSTRACT:

Background

Recombinant mouse GH (mGH) is a critical tool for investigating GH-GH receptor (GHR) interactions in rodent models. However, while numerous methods exist for producing human GH, detailed protocols for the expression and purification of recombinant mGH remain scarce in the literature.

Methods

We developed a method for refolding mGH from inclusion bodies produced in Escherichia coli (E. coli). Recombinant mGH was fused with a N-terminal thioredoxin (Trx) tag and was expressed as inclusion body proteins in E. coli when induced at 30 °C. Inclusion bodies were isolated and solubilised under mild conditions (50 mM Tris-HCl, 2 M urea, pH 10.5) and freeze-thawed.

Results

Three rounds of freeze-thawing improved solubilisation when compared to a single round. Subsequ

SUBMITTER: Kim M 

PROVIDER: S-EPMC12185587 | biostudies-literature | 2025 Jun

REPOSITORIES: biostudies-literature