A novel method for the large-scale production of PG-CNP37, a C-type natriuretic peptide analogue

Achondroplasia is the most common form of human dwarfism caused by a mutation in the fibroblast growth factor receptor 3 (FGFR3), resulting in abnormal endochondral bone formation. C-type natriuretic peptide (CNP) is a potent stimulator of endochondral bone growth and represents a potential therapy for achondroplasia. We have developed a novel, simple and cost effective method to produce a CNP analogue, PG-CNP37, at a large scale from Escherichia coli. A PG-CNP37 fusion protein was over-expressed as inclusion bodies in E. coli, which were purified then cleaved by formic acid to release the PG-CNP37 peptide. Approximately 0.5 g of 95% pure, soluble and active PG-CNP37 peptide was produced from 1 L of culture using this method and may represent a viable means for large-scale production of other therapeutic peptides.

► Novel method for producing an active 39-amino acid CNP analogue, PG-CNP37. ► Overexpression as inclusion bodies followed by formic acid cleavage. ► Achieved 80% pure PG-CNP37 peptide by a neutralization and centrifugation step. ► Final yield of 0.5 g/L PG-CNP37 peptide with 95% purity.