NON-CYTOLYTIC EXTRACTION OF MURINE TUMOR-SPECIFIC TRANSPLANTATION ANTIGENS WITH THE NON-IONIC DETERGENT OCTYL-BETA-DEUTERIUM-GLUCOPYRANOSIDE

  • 1 January 1982
    • journal article
    • research article
    • Vol. 69  (1) , 131-136
Abstract
The nonionic detergent octyl-.beta.-D-glucopyranoside (C8Glu) was used to extract immunogenic tumor-specific transplantation antigen (TSTA) from intact cells and purified plasma membranes of the 3-methylcholanthrene-induced mouse fibrosarcoma MCA-F. Pretreatment of syngeneic C3H/HeJ mice with 100 .mu.g of C8Glu extracts induced specific immunoprotection such that mice resisted the outgrowth of MCA-F but not the antigenically distinct tumors MCA-D or MCA-2A. Incubation of intact cells with 7 mM (0.2%) C8Glu for 30 min at 23.degree. C was judged to be noncytolytic because extracted cells excluded trypan blue. Preparative isoelectric focusing partially purified the MCA-F specific antigen from crude C8Glu extracts into the pH 6.5-6.8 region of the gradient. Electrofocusing yielded 60% of the applied antigen activity and a 4- to 5-fold increase in specific activity. In addition, immunoprotective activity was obtained in 2% C8Glu extracts of MCA-F plasma membranes, confirming the membrane localization of the MCA-F TSTA. Three properties of C8Glu rendered it an attractive agent for the preparation of cell surface proteins: a nonionic character, large critical micellar concentration, and its capacity to extract antigens without complete membrane solubilization or cell disruption.

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