Poly(amidoamine)s (PAAs) are water-soluble synthetic polymers designed to be biodegradable and biocompatible. Moreover, they display membrane disruptive properties in response to a decrease in pH. This attribute confers PAAs with endosomolytic properties in vitro and in vivo. A model system was developed to quantify their ability to promote the endosomal escape of macromolecules that may be interesting as therapeutic agents. Here, two PAAs (ISA 1 and 4) were incubated with B16F10 cells in vitro together with two non-permeant toxins: either ricin A-chain (RTA) or gelonin. The relatively non-toxic PAAs ISA 1 and 4 (IC 50>1.5 mg/ml) restored activity to the inherently inert toxins. The IC 50 values for the ISA 1/RTA and ISA 1/gelonin combinations were 0.65±0.05 and 0.55±0.12 mg/ml, respectively. Similarly, when ISA 4 was incubated with a non-toxic combination of RTA and gelonin the IC 50 value decreased to 0.57±0.03 and 0.43±0.26 mg/ml, respectively. In contrast, the neutral polymer dextran and the PAA ISA 22 were unable to mediate this effect. These observations suggest that specific PAA–toxin combinations warrant further development as novel therapeutics.