AN ASOLECTIN ADSORBED SUBSTRATE FOR PROACCELERIN ASSAY

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Abstract
Asolectin in the concentration of 25 mg/ ml of normal citrated plasma is capable of specifically binding proaccelerin. Sedimentation of the asolectin-proaccelerin complex after incubation (60 minutes 37[degree]C) of a Potter homogenized suspension of asolectin (25 mg/ml of plasma) in plasma diluted 1:6 with distilled water is effected by spinning at 40,000 rpm in a Spinco preparatory centrifuge for 1 hour (4[degree]C). The clear supernate is aspirated, pooled, quickly shell-frozen, lyophilized, and reconstituted to the original plasma volume with distilled water. pH is then corrected to 7.3 and aliquots frozen at minus 20[degree]C for use as substrate in a one-stage system for proaccelerin determination. Mutual failure of this proaccelerin deficient plasma and para-hemophilia plasma to correct one another was shown. Control plasma similarly diluted, alkalinized to pH 9.0, incubated, centrifuged, lyophilized, reconstituted and pH readjusted to 7.3 showed no comparable loss of proaccelerin. Evidence of the specificity of the substrate for proaccelerin activity and its insensitivity to proconvertin and prothrombin concentrations in the test plasma was the finding of a constant proaccelerin content in varying percentage mixtures of BaSO4 adsorbed normal plasma and the same oxalated normal plasma unadsorbed.