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Nucleic Acids Research, 2003, Vol. 31, No. 2 e4
© 2003 Oxford University Press

A rapid biosensor chip assay for measuring of telomerase activity using surface plasmon resonance

Chihaya Maesawa*, Toru Inaba1, Hidetoshi Sato, Sin Iijima1, Kaoru Ishida1, Masanori Terashima2, Ryo Sato, Michihiro Suzuki, Akiko Yashima, Satoshi Ogasawara1, Hiroki Oikawa, Nobuhiro Sato1, Kazuyoshi Saito1 and Tomoyuki Masuda

Department of Pathology and 1 Department of Surgery, Iwate Medical University School of Medicine, Uchimaru 19-1, Morioka 202-8505, Japan and 2 Department of Surgery 1, Fukushima Medical University, Fukushima 960-1295, Japan

*To whom correspondence should be addressed. Tel: +81 19 651 5111; Fax: +81 19 629 9340; Email: chihaya{at}iwate-med.ac.jp

Considerable interest has been focused on telomerase because of its potential use in assays for cancer diagnosis, and for anti-telomerase drugs as a strategy for cancer chemotherapy. A number of assays based on the polymerase chain reaction (PCR) have been developed for evaluation of telomerase activity. To overcome the disadvantages of the conventional telomerase assay [telomeric repeat amplification protocol (TRAP)] related to PCR artifacts and troublesome post-PCR procedures, we have developed a telomeric repeat elongation (TRE) assay which directly measures telomerase activity as the telomeric elongation rate by biosensor technology using surface plasmon resonance (SPR). 5'-Biotinylated oligomers containing telomeric repeats were immobilized on streptavidin-pretreated dextran sensor surfaces in situ using the BIACORE apparatus. Subsequently, the oligomers associated with the telomerase extracts were elongated in the BIACORE apparatus. The rate of TRE was calculated by measuring the SPR signals. We examined elongation rates by the TRE assay in 18 cancer and three normal human fibroblast cell lines, and 12 human primary carcinomas and matching normal tissues. The elongation rates increased in a concentration- and time-dependent manner. Those of cancer cells were two to 10 times higher than fibroblast cell lines and normal tissues. Telomerase activities and its inhibitory effects of anti-telomerase agents as measured by both the TRE and TRAP assays showed a good correlation. Our assay allows precise quantitative comparison of a wide range of human cells from somatic cells to carcinoma cells. TRE assay is suitable for practical use in the assessment of telomerase activity in preclinical and clinical trials of telomerase-based therapies, because of its reproducibility, rapidity and simplicity.


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