Expression and purification of biologically active v-sis/platelet-derived growth factor B protein by using a baculovirus vector system.

Malignant transformation induced by simian sarcoma virus is mediated by its v-sis protein, the monkey homolog of the platelet-derived growth factor (PDGF) B chain. By use of an appropriately engineered baculovirus expression vector, the v-sis protein was expressed in the insect cell line Spodoptera frugiperda (Sf9) at a level 50- to 100-fold higher than that observed with overexpression in mammalian-cell transfectants. The sis protein produced by Sf9 cells underwent processing similar to that observed in mammalian cells, including efficient disulfide-linked dimer formation. Moreover, the recombinant sis protein was capable of binding PDGF receptors and inducing DNA synthesis as efficiently as PDGF-B synthesized by mammalian cells. A significant fraction of sis protein was released from Sf9 cells, which made possible a one-step immunoaffinity purification to near homogeneity with a 40% recovery of biological activity. These results demonstrate that a protein whose normal processing requires both intrachain and interchain disulfide-bridge formation can be efficiently expressed in a biologically active form in insect cells by using a baculovirus vector system.