Preliminary studies with L-asparaginase bound to implantable bovine collagen heterografts: a potential long-term, sustained dosage, antitumor enzyme therapy system.
In this study, L-Asparaginase has been bound to collagen heterografts derived from carotid bovine arteries. The immobilization procedure utilizes both non-covalent and covalent interactions to fix the enzyme. Binding of the enzyme to the graft material was shown to the pH dependent, with optimum binding occurring at pH 6.0 and pH 8.5. Amidohydrolysis by the bound enzyme exhibited zero-order kinetic behavior at substrate saturating conditions. Total apparent asparaginase activity expressed by the grafts as a function of the number of repeated in vitro assay trials demonstrated that over a span of 3 months of intermittent storage and use, the enzyme-grafts retained as much as 62% of their initial activities. Implantation of 4 asparaginase-collagen grafts in various locations of the thoracic and abdominal aorta resulting in prolonged reductions of plasma asparagine levels in 3 of the 4 implants. Presence of plasma asparaginase was checked in one of the four implants and determined to be less than 2 X 10(-4) I.U./ml. Removal of grafts from 3 of the 4 animal subjects showed reductions in the apparent asparaginase activity expressed by the grafts of 7 to 70 percent after in vivo contact times which varied from 6 to 15 days.
Preliminary studies with L-asparaginase bound to implantable bovine collagen heterografts: a potential long-term, sustained dosage, antitumor enzyme therapy system. In this study, L-Asparaginase has been bound to collagen heterografts derived from carotid bovine arteries. The immobilization procedure utilizes both non-covalent and covalent interactions to fix the enzyme. Binding of the enzyme to the graft material was shown to the pH dependent, with optimum binding occurring at pH 6.0 and pH 8.5. Amidohydrolysis by the bound enzyme exhibited zero-order kinetic behavior at substrate saturating conditions. Total apparent asparaginase activity expressed by the grafts as a function of the number of repeated in vitro assay trials demonstrated that over a span of 3 months of intermittent storage and use, the enzyme-grafts retained as much as 62% of their initial activities. Implantation of 4 asparaginase-collagen grafts in various locations of the thoracic and abdominal aorta resulting in prolonged reductions of plasma asparagine levels in 3 of the 4 implants. Presence of plasma asparaginase was checked in one of the four implants and determined to be less than 2 X 10(-4) I.U./ml. Removal of grafts from 3 of the 4 animal subjects showed reductions in the apparent asparaginase activity expressed by the grafts of 7 to 70 percent after in vivo contact times which varied from 6 to 15 days.