Purification of immunoprotective tumor antigens by preparative isotachophoresis.
Tumor-specific transplantation antigens (TSTA) were purified from 3 M KCl and butanol extracts of C3H/HeJ 3-methylcholanthrene-induced fibrosarcomas by preparative isotachophoresis (pITP). Fractions from pITP which reacted with antisera to TSTA preparations in an enzyme-linked immunospecific assay were tested in vivo for induction of resistance to the growth of transplanted tumor cells. Isotachophoresis of crude 3 M KCl extracts yielded TSTA that was immunogenic at doses between 17 and 124 micrograms. Isotachophoresis of TSTA partially purified from crude 3 M KCl or butanol extracts by preparative isoelectric focusing (pIEF) of 3 M KCl and butanol extracts yielded TSTA that was immunogenic over a two-fold log dose range. As little as 10 ng purified TSTA reduced tumor growth by 50%. Tumor growth reduction was specific, and immunized animals survived longer than non-immunized controls. A purification of 10,000-fold over crude 3 M KCl extracts and 2,500-fold over crude butanol extracts was obtained. These results suggest that TSTA from murine tumor cells is preparatively purified by following extraction with pIEF and then pITP.
D000596:Amino Acids; D000665:Ampholyte Mixtures; D000818:Animals; D000951:Antigens, Neoplasm / Q000008:administration & dosage / Q000276:immunology / Q000302:isolation & purification*; D000440:Butanols; D002457:Cell Extracts / Q000032:analysis / Q000276:immunology; D002471:Cell Transformation, Neoplastic / Q000032:analysis / Q000276:immunology; D005260:Female; D005354:Fibrosarcoma / Q000032:analysis / Q000276:immunology* / Q000628:therapy; D006649:Histocompatibility Antigens / Q000008:administration & dosage / Q000276:immunology / Q000302:isolation & purification*; D007167:Immunotherapy; D007525:Isoelectric Focusing / Q000379:methods*; D051379:Mice; D008809:Mice, Inbred C3H; D008817:Mice, Mutant Strains; D011189:Potassium Chloride; D011817:Rabbits
D000596:Amino Acids; D000665:Ampholyte Mixtures; D000951:Antigens, Neoplasm; D000440:Butanols; D002457:Cell Extracts; D006649:Histocompatibility Antigens; C029237:tumor-associated transplantation antigen; D011189:Potassium Chloride