Novel recombinant protein constructs for improved insulin-like growth factor-1 delivery

Abstract

Four novel recombinant protein constructs were designed to improve the delivery of insulin-like growth factor-1 (IGF-1). The IGF-1 sequence was subcloned into a HisMax plasmid and expressed at low levels by transient transfection in 293 cells. Site-specific mutagenesis was used to insert all four construct sequences into the IGF-1 HisMax plasmid, and to insert a secretory sequence into all four construct plasmids. A stable cell line selection in 293 and CHO cells using Zeocin was attempted. The four construct sequences were then subcloned from the HisMax plasmids to TrcHis plasmids and expressed in bacteria for higher production efficiency. Ni-NTA purified protein was detected in three of the four constructs. These proteins may ultimately be useful for myocardial delivery of IGF-1 to facilitate cardiac repair.