A DEAE-cellulose batch technique for antilymphyocyte globulin preparation.

Absorption: With the original technique (1) the horse serum decomplemented by heating to 56 0 C. and then absorbed against cells and pooled serum of the species to be treated; tion was done first to avoid hemolysis of the added red cedure had two flaws. First, the use of serum (rather than p.,,"~ ... aJ ICtJ,:'lgror absorption precluded the removal of antifibrinogen and bocyte antibodies. Secondly, the removal of horse antibodies -O----~~.~,I5j,lJu.LJ other human plasma proteins was made inefficient by the abi!eni:.e;l1ilirevi of complement at this stage. These objections were eliminated minimized. Whole blood was collected from multiple donors ethylene diamine tetra-acetic acid (EDTA) disodium as an coagulant in order to prevent platelet aggregation. The red plasma, and platelets were separated with differential The human (or dog) plasma was added to the unaltered horse and incubated for 2 hr. at 37 0 C. and 18 hr. at 40 C. vVith thet: .~:~q~~.~,~ tion of one part human or dog plasma to 10 parts of horse "'f most of the horse precipitating antibodies were eliminated. this incubation, the platelets isolated earlier were added to the ture in quantities determined by the premeasured titers of the platelet activity in the ALS and by standard curves showing the ber of platelets needed at various titer levels. Platelet absorption not reduce the antiwhite cell titer. The supernatant was now complemented by heating at 56 0 C. for 30 min. and absorbed red cells; hemolysis did not occur. The final product had no