AN ORTHOGONAL PROCESS FOR PURIFICATION OF RECOMBINANT HUMAN PARATHYROID HORMONE (RHPTH) (1-34)

摘要

The present invention discloses a process for the preparation of rhPTH (1-34) also known as teriparatide by construction of a novel nucleotide, as an Ncol IXhoI fragment as set forth in SEQ.ID No.:l encoding a chimeric fusion protein as set forth in SEQ.ID No.:2 comprising of a fusion partner consisting of 41 amino acids belonging to Escherichia coli P-galactosidase (LacZ) gene, an endoeptidase cleavage site, rhPTH (1-34) gene fragment, cloning the said nucleotide in an expression vector under the control of T7 promoter, transforming Escherichia coli with the said vector and expressing the chimeric fusion protein in fed batch fermentation. The present invention further discloses a low feed rate lactose induction for optimized expression of rhPTH (1-34) in Escherichia coli. The present invention also discloses an unique, novel two step orthogonal purification process for rhPTH (1-34) comprising of cation exchange chromatography optionally followed by preparative chromatography selected from HIC or RP-HPLC to yield a target protein of 99% purity. The present invention discloses a simple, cost-effective, environmentally benign method of producing high purity rhPTH(l-34).