Purification of human erythrocyte phosphoglycerate kinase by dye ligand affinity chromatography.

A new method for the purification of human erythrocyte phosphoglycerate-kinase involving affinity chromatography on dye-ligand media (Red A), in the presence of 3-phosphoglycerate and ATP, is described. The method is rapid and technically simple. The purity of the enzyme was verified by electrophoresis in polyacrylamide gel in the presence of sodium dodecylsulphate, by amino acid analysis and by immunoprecipitation in Ouchterlony plates.

Isoelectric focusing of human hemoglobins.

Isoelectric focusing on thin layer of acrylamide gel has been used for the characterization of 79 different variants of hemoglobin A. This method has replaced the cellulose acetate electrophoresis in clinical laboratories in Martinique, Guadeloupe and Creteil for the detection of abnormal hemoglobins in populations at risk. Up to now 15,000 samples from adults have been evaluated.

Hemoglobin Bougardirey-Mali beta 119 (GH2) Gly replaced by Val. An electrophoretically silent variant migrating in isoelectrofocusing as Hb F.

Hemoglobin Bougardirey-Mali was detected by isoelectrofocusing during a screening in a 32 years old African, a native of Mali. This abnormal Hb, representing 35% of the total, exhibited the same pI as that of Hb F. In contrast, it was indistinguishable from Hb A in all the electrophoretic systems tested, and equally by its resistance to alkaline denaturation.

Delayed type hypersensitivity to bovine serum albumin and to lipid-conjugated bovine serum albumin in mice.

Delayed type hypersensitivity (DTH) to bovine serum albumin (BSA) and to lipid-conjugated BSA were studied comparatively. Unlike the case of BSA with which no DTH can be detected with native antigen, injection of butyric-conjugated BSA (Bu-BSA) in sensitized mice provokes a typical DTH for an early and limited period. Alum-precipitated Bu-BSA (Al-Bu-BSA) provokes from the beginning a stronger DTH which persists a much longer period.