Isolation of plasma-membrane components from cultured human pancreatic cancer cells by immuno-affinity chromatography of anti-beta 2M sepharose 6MB.
Human pancreatic exocrine adenocarcinoma cells established in tissue culture expressed both HLA and beta 2-microglobulin (beta 2M). Plasma-membrane components of this pancreatic cancer cell line were purified from plasma membrane fractions enriched by sucrose density-gradient centrifugation, using immunoaffinity chromatography on immobilized anti-human beta 2M antibody. Both rabbit and mouse monoclonal anti-beta 2M IgG were used, with a 20--25-fold overall purification of 5'-nucleotidase. The method was applicable to 5 x 10(7) cells and permitted the solubilization of membranes retained on the column, with the selective desorption of components not associated with beta 2M before the subsequent elution at pH 3 of beta 2M-associated macromolecules. The acid eluate contained one major and two minor bands in the 40--45,000 mol.-wt range with two additional enriched components of 18,000 and 22,000 dalton. A major carbohydrate-containing component of high mol. wt was also found to be associated with the pancreatic cancer-cell plasma membrane.
D000230:Adenocarcinoma / Q000276:immunology*; D000906:Antibodies; D001609:Beta-Globulins / Q000276:immunology*; D002458:Cell Fractionation; D002462:Cell Membrane / Q000276:immunology*; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D006680:HLA Antigens; D006801:Humans; D008565:Membrane Proteins / Q000276:immunology; D009363:Neoplasm Proteins / Q000276:immunology; D010190:Pancreatic Neoplasms / Q000276:immunology*; D001613:beta 2-Microglobulin / Q000276:immunology*
D000906:Antibodies; D001609:Beta-Globulins; D006680:HLA Antigens; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D001613:beta 2-Microglobulin