Methods for Thymidylate Synthase Pharmacodynamics: Serial Biopsy, Free and Total TS, FdUMP and dUMP, and H4PteGlu and CH2-H4PteGlu Assays
作者： Colin Paul Spears , Bengt G. Gustavsson
DOI: 10.1007/978-1-4684-5607-3_9
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摘要: This report details our methods for performance of the major parameters related to quantitation TS inhibition resulting from fluoropyrimidine administration patients, equally applicable preclinical studies. Sampling tumors before and after drug treatment is done by 4 mm disposable punch biopsy or forceps via subcutaneous tunneling. Homogenates are prepared using N2 polytron-mincing. Cytosolic free measured either tritium-release method small biopsies [3H]FdUMP ligand-binding. FdUMP dUMP separated DEAE-cellulose column competitive binding [14C]dTMP synthesis Moran methods. Total, postFUra pre-incubation dissociation FdUMP-bound neutral charcoal removal cytosolic ligands. H4PteGlu CH2-H4PteGlu Priest L. Casei TS. The materials described in sufficient detail permit wide application this approach.
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