The Synthesis of L-tryptophan Degrading Bioreactors

Abstract

To design a bioreactor for removing the potential cancer nutrient L-tryptophan from blood, the L-tryptophan degrading enzyme tryptophan side chain oxydase (TSO) was chemically bound to glutaraldehyde activated gamma amino silane silica and to Zetaffinity microcolumns consisting of a glutaraldehyde activated polyacrylic-cellulose copolymer. Five experiments were carried out in sheep and six experiments in rabbits using a closed circuit plasmapheresis bioreactor system. L-tryptophan in sheep was degraded by the silica bioreactor in a single pass to undetectable levels as measured by high performance liquid chromatography (HPLC). Zetaffinity bioreactors degraded L-tryptophan in rabbits to more than 95% in a single pass. Whole blood L-tryptophan levels changed little throughout the experiment indicating a vast extravascular tryptophan pool. Enzyme leakage from the bioreactor was less than 10⁻⁵ IU TSO per ml plasma. The procedures were tolerated well by the animals without any change in vital signs