Expression of a recombinant human growth hormone binding protein in baculovirus/insect cell system

摘要

An eucaryotic recombinant human growth hormone binding protein (rGHBP) was expressed in baculovirus-infeeted insect cells and purified by affinity chromatography from culture supernatant. This mannose-rich 34-kDa protein specifically bound human growth hormone (hGH) with the same affinity (kDa = 0.42 x 10~(-9) M) than the 51.5 kDa GHBP we purified and characterised from human plasma (kDa = 1.1 x 10~(-9) M). A high molecular form of the rGHBP was detected by silver-stained SDS-PAGE, Western blot (mAb 263), affinity cross-linking and Western ligand blot with ~(125)I-hGH. Reduction experiments with β-mercaptoethanol suggested that this form involved a disulfide bound between two rGHBPs.