Isolation of a papain-solubilised major transplantation antigen from rat leukaemic cells.
The major transplantation antigen of the PVG rat, Rt-1c, has been isolated from PVG leukaemic cells in milligram quantities for use in biological experiments related to transplantation. A 3-stage isolation procedure was developed involving the preparation of plasma membranes, digestion of the membranes with papain to release an active fragment of the Rt-1c antigen and fractionation of the digest on Sephacryl columns to give an active product. Activity was followed by a newly developed inhibition assay in which extracts of cells were incubated with a reference anti-PVG antiserum, excess unreacted antibody being quantitated by radioactive binding to PVG leukaemic cells glutaraldehyde-fixed to flexible microtitre plates. In addition to Rt-1c the preparation contained a second glycoprotein component. The preparation shows allelic specificity in that it inhibits the binding of anti-PVG antiserum to PVG cells, but not the binding of anti-DA serum to DA cells. The preparation represents a 16% yield of the activity of the cells from which it was derived.
D000818:Animals; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006649:Histocompatibility Antigens / Q000302:isolation & purification*; D007942:Leukemia, Experimental / Q000276:immunology*; D008198:Lymph Nodes / Q000378:metabolism; D010206:Papain / Q000494:pharmacology*; D051381:Rats; D011914:Rats, Inbred BN; D012995:Solubility
D006649:Histocompatibility Antigens; D010206:Papain