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Methods for Thymidylate Synthase Pharmacodynamics: Serial Biopsy, Free and Total TS, FdUMP and dUMP, and H4PteGlu and CH2-H4PteGlu Assays

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The Expanding Role of Folates and Fluoropyrimidines in Cancer Chemotherapy

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 244))

Abstract

This report details our methods for performance of the major parameters related to quantitation of TS inhibition resulting from fluoropyrimidine administration to patients, methods equally applicable to preclinical studies. Sampling of tumors before and after drug treatment is done by 4 mm disposable punch biopsy or forceps biopsy via subcutaneous tunneling. Homogenates are prepared using N2 or polytron-mincing. Cytosolic free TS is measured by either the tritium-release method for small biopsies or by [3H]FdUMP ligand-binding. FdUMP and dUMP are separated by DEAE-cellulose column and measured by competitive binding and [14C]dTMP synthesis by the Moran methods. Total, postFUra TS is measured by pre-incubation dissociation of FdUMP-bound TS after neutral charcoal removal of cytosolic ligands. H4PteGlu and CH2-H4PteGlu are measured by the Priest method using L. Casei TS. The materials and methods are described in sufficient detail to permit wide application of this approach.

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References

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Authors and Affiliations

  1. Cancer Research Laboratory, University of Southern California Comprehensive Cancer Center, 1303 N. Mission Road, Los Angeles, CA, 90033, USA

    Colin Paul Spears & Bengt G. Gustavsson

  2. Department of Surgery Östra Hospital, University of Göteborg, Göteborg, Sweden

    Colin Paul Spears & Bengt G. Gustavsson

Authors
  1. Colin Paul Spears
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  2. Bengt G. Gustavsson
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Editors and Affiliations

  1. Roswell Park Memorial Institute, Buffalo, New York, USA

    Youcef Rustum  & John J. McGuire  & 

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© 1988 Plenum Press, New York

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Spears, C.P., Gustavsson, B.G. (1988). Methods for Thymidylate Synthase Pharmacodynamics: Serial Biopsy, Free and Total TS, FdUMP and dUMP, and H4PteGlu and CH2-H4PteGlu Assays. In: Rustum, Y., McGuire, J.J. (eds) The Expanding Role of Folates and Fluoropyrimidines in Cancer Chemotherapy. Advances in Experimental Medicine and Biology, vol 244. Springer, New York, NY. https://doi.org/10.1007/978-1-4684-5607-3_9

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  • DOI: https://doi.org/10.1007/978-1-4684-5607-3_9

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  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-4684-5609-7

  • Online ISBN: 978-1-4684-5607-3

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