Immunoadsorbent Chromatography of Hypoxanthine-Guanine Phosphoribosyltransferase

Abstract

Recent evidence indicates that the virtual absence of hypoxanthine-guanine phosphoribosyltransferase (HGPRT) in patients with the Lesch-Nyhan syndrome is due in most if not all instances to a mutation(s) on the gene coding for the HGPRT protein (Kelley and Meade, 1971; Rubin, et al., 1971; Arnold, Meade and Kelley, 1972). This mutation(s) results in the synthesis of a normal amount of a catalytically defective yet immunoreactive HGPRT enzyme protein. We have purified normal human HGPRT from erythrocytes and have prepared a highly specific antiserum in rabbits (anti-HGPRT) which displays a single precipitin line on immunodiffusion and immunoelectrophoresis with both normal hemolysate and hemolysate from patients with the Lesch-Nyhan syndrome. We have succeeded in coupling a partially-purified preparation of anti-HGPRT to cyanogen bromide activated sepharose and will present data which indicates that this technique is a promising method for the rapid isolation of both normal and mutant HGPRT.