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Purification of immunoprotective tumor antigens by preparative isotachophoresis

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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 μg. 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.

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Authors and Affiliations

  1. The University of Texas Health Science Center Medical School and Graduate School of Biomedical Sciences, 77030, Houston, TX, USA

    Thomas L. Saunders, Barry D. Kahan & Neal R. Pellis

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  1. Thomas L. Saunders
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  2. Barry D. Kahan
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  3. Neal R. Pellis
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Saunders, T.L., Kahan, B.D. & Pellis, N.R. Purification of immunoprotective tumor antigens by preparative isotachophoresis. Cancer Immunol Immunother 16, 101–108 (1983). https://doi.org/10.1007/BF00199240

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  • DOI: https://doi.org/10.1007/BF00199240

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