Detection of human hematopoietic differentiationrelated glycoproteins: The Western enzyme-linked lectin analysis

A technique is introduced (Western enzyme-linked lectin analysis, WELLA) for detecting lectin-reactive cellular glycoproteins after separation on the basis of molecular weight in sodium dodecyl sulfate (SDS) polyacrylamide gels. Lectin-reactive glycoproteins are detected on Western transfers by reaction with lectin-peroxidase conjugates followed by development with hydrogen, peroxide and 4-chloro-1-naphthol which forms a purple-gray precipitate. WELLA is more rapid, more sensitive, and the bands are highly reproducible and better resolved than those obtained, by autoradiography of fluorography.Using this technique, we have detected human differentiation-related glycoproteins on cells of different hematological lineages. Both wheat germ agglutinin-peroxidase (WGA-P) and concanavalin A-peroxidase (ConA-P) detected distinct glycoprotein patterns on isolated peripheral blood platelets, lymphocytes, monocytes, erythrocytes and granulocytes. WGA-P detected numerous similarities between immature myeloid cells isolated from bone marrow and acute myelogenous leukemia cells, including major glycoproteins at 20 and 25 kDa. ConA-P detected a similar pattern of glycoproteins between isolated peripheral blood lymphocytes and T-cell acute lymphoblastic leukemia (T-ALL) cells. The T-ALL cells, however, had a major 200 kDa glycoprotein not present on lymphocytes. WGA-P also showed nearly identical patterns between the lymphocytes and the T-ALL cells, but detected prominent 200 and 250 kDa glycoproteins on the T-ALL cells which were absent from the lymphocytes. We have also detected polymorphic differences in the glycoproteins on lymphocytes from normal donors in the range of 95-100 kDa using ConA-P.Abbreviations WELLA Western enzyme-linked lectin analysis - SDS sodium dodecyl sulfate - BSA bovine serum albumin - PVP polyvinylpyrrolidone - PBS phosphate-buffered saline - AML acute myelogenous leukemia - ALL acute lymphocytic leukemia - WGA wheat germ agglutinin - Con A concanavalin A - WGA-P wheat germ agglutinin-peroxidase conjugate - ConA-P concanavalin A-peroxidase conjugate