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 w...

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Bibliographic Details
Published in:Cancer Immunology Immunotherapy 1983-01, Vol.16 (2), p.101-108
Main Authors: SAUNDERS, T. L, KAHAN, B. D, PELLIS, N. R
Format: Article
Language:English
Subjects:
Amino Acids
Ampholyte Mixtures
Animals
antigen (tumor-specific)
Antigens, Neoplasm - administration & dosage
Antigens, Neoplasm - immunology
Antigens, Neoplasm - isolation & purification
Biological and medical sciences
Butanols
Cell Extracts - analysis
Cell Extracts - immunology
Cell Transformation, Neoplastic - analysis
Cell Transformation, Neoplastic - immunology
Female
fibrosarcoma
Fibrosarcoma - analysis
Fibrosarcoma - immunology
Fibrosarcoma - therapy
Histocompatibility Antigens - administration & dosage
Histocompatibility Antigens - immunology
Histocompatibility Antigens - isolation & purification
Host-tumor relations. Immunology. Biological markers
Immunotherapy
Isoelectric Focusing - methods
isotachophoresis
Medical sciences
Mice
Mice, Inbred C3H
Mice, Mutant Strains
Original
Potassium Chloride
Rabbits
Tumors
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Summary: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.
ISSN:0340-7004
1432-0851
DOI:10.1007/BF00199240