Sensitive nonradiometric method for determining thymidine kinase 1 activity

Clinical Chemistry
Anders OhrvikStaffan Eriksson

Abstract

Thymidine kinase 1 (TK1) is a cytoplasmic enzyme, produced only in the S-phase of proliferating cells, that has potential as a tumor marker. Specific determination of TK1 in serum is difficult, in part because of differences in the physical properties of serum TK1 compared with cytoplasmic TK1. The first step in the new assay was phosphorylation of 3'-azido-2',3'-deoxythymidine (AZT) to AZT 5'-monophosphate (AZTMP) by TK1 present in patient material. The AZTMP formed was measured in a competitive immunoassay with specific anti-AZTMP antibodies and AZTMP-labeled peroxidase. Results were compared with those of a TK radioenzyme assay (REA) for 78 samples from patients suffering from hematologic diseases. The detection limit was 78 microIU/L, and within-run CVs <20% were seen for samples with TK1 down to 130 microIU/L. Cross-determination of the mitochondrial isoenzyme TK2 activity was <0.1%. Between-assay imprecision (CV) was 3.5-7.4%, and the within-assay imprecision was 4.1-9.1%. In studies of recovery and linearity on dilution, measured values ranged from 84% to 115% of expected at concentrations of 0.26-10.4 mIU/L. Results of the new assay (mIU/L) = 0.109 x TK REA (U/L) + 0.092. Heterophilic antibodies did not interfere in the...Continue Reading

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Citations

Feb 1, 2008·Expert Opinion on Medical Diagnostics·Ondrej Topolcan, Lubos Holubec
Aug 30, 2008·Journal of Cellular and Molecular Medicine·Ossie F DysonShaw M Akula
Feb 10, 2011·Veterinary and Comparative Oncology·H von Euler, S Eriksson
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