Cytotoxic Activity of Recombinant bFGF–rViscumin Fusion Proteins

Abstract A fusion protein (bFGF–rMLA), containing the mitogen basic fibroblast growth factor (bFGF) and the cytotoxic component of rViscumin (recombinant mistletoe lectin), the enzymatic A-chain (rMLA), was expressed in Escherichia coli, purified, and functionally characterized. bFGF–rMLA is cytotoxic for mouse B16 melanoma cells expressing the FGF receptor with an IC 50 value of approximately 1 nM. rMLA shows no significant effect on the viability of the B16 cells up to a concentration of 141 nM. Additionally, bFGF–rMLA was associated with the rViscumin B-chain (rMLB) in an in vitro folding procedure. The IC 50 value of bFGF–rMLA/rMLB to B16 cells in the presence of lactose—to block rMLB lectin activity—was 134 pM. Thus, it was possible to enhance the efficacy of a rViscumin A-chain mitotoxin through addition of rMLB. We conclude that rViscumin fusion proteins may be generally applicable for the receptor-specific inactivation of target cells and point out their potential in drug development.