Jump to ContentJump to Main Navigation
Show Summary Details
More options …

Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

Editorial Board: Buchner, Johannes / Lei, Ming / Ludwig, Stephan / Thomas, Douglas D. / Turk, Boris / Wittinghofer, Alfred


IMPACT FACTOR 2018: 3.014
5-year IMPACT FACTOR: 3.162

CiteScore 2018: 3.09

SCImago Journal Rank (SJR) 2018: 1.482
Source Normalized Impact per Paper (SNIP) 2018: 0.820

Online
ISSN
1437-4315
See all formats and pricing
More options …
Volume 383, Issue 10

Issues

Real-Time Determination of Telomerase Activity in Cell Extracts Using an Optical Biosensor

P. M. Schmidt / E. Matthes / F. W. Scheller / M. Bienert / C. Lehmann / A. Ehrlich / F. F. Bier
Published Online: 2005-06-01 | DOI: https://doi.org/10.1515/BC.2002.186

Abstract

A biosensoric approach has been developed to determine the activity of telomerase in tumor cell lysates. An optical sensor, the grating coupler, was used to monitor the association and dissociation of unlabeled compounds on the sensor surface in real time, by virtue of an evanescent field. An oligonucleotide was immobilized on the surface of the optical biosensor and linked with two other oligonucleotides by complementary sequences in an overlapping manner. The 3end of the last one carried the sequence of the telomeric substrate (TS) primer used for elongation by telomerase in the telomeric repeat amplification protocol (TRAP) assay. This primer sequence was phosphorothioate (PS) modified, which is known to strongly increase the affinity to the primer binding site of telomerase protein and consequently the velocity of the telomerase reaction. We show that the PS primer binds to the modified biosensor and is elongated effectively by the telomerase from HL-60 cell lysates. A synthesis rate of 1 nucleotide/min was determined. The inhibitory effect of peptide nucleic acid (PNA) was shown by using immobilized TS. The velocity of the telomerase reaction was slowed down and the signal intensity was below the signaltonoise ratio. Most nucleic acid detection systems use amplification steps such as polymerase chain reaction (PCR) to increase the amount of the probe. Since telomerase is a polymerase itself amplification of DNA by PCR is not required. Furthermore, no purification steps were required since all measurements were performed with crude cell extract.

About the article

Published Online: 2005-06-01

Published in Print: 2002-10-06


Citation Information: Biological Chemistry, Volume 383, Issue 10, Pages 1659–1666, ISSN (Print) 1431-6730, DOI: https://doi.org/10.1515/BC.2002.186.

Export Citation

Citing Articles

Here you can find all Crossref-listed publications in which this article is cited. If you would like to receive automatic email messages as soon as this article is cited in other publications, simply activate the “Citation Alert” on the top of this page.

[1]
Peter M Schmidt, Christine Lehmann, Eckart Matthes, and Frank F Bier
Biosensors and Bioelectronics, 2002, Volume 17, Number 11-12, Page 1081
[3]
Xiafei Zhang, Rui Cheng, Zhilu Shi, and Yan Jin
Biosensors and Bioelectronics, 2016, Volume 75, Page 101
[4]
Leilei Tian and Yossi Weizmann
Journal of the American Chemical Society, 2013, Volume 135, Number 5, Page 1661
[5]
Xi Zhu, Huifeng Xu, Ruolan Lin, Guidi Yang, Zhenyu Lin, and Guonan Chen
Chemical Communications, 2014, Volume 50, Number 58, Page 7897
[6]
Andrew T. Ludlow, Jerome D. Robin, Mohammed Sayed, Claudia M. Litterst, Dawne N. Shelton, Jerry W. Shay, and Woodring E. Wright
Nucleic Acids Research, 2014, Volume 42, Number 13, Page e104
[7]
Eliona Kulla and Evgeny Katz
Sensors, 2008, Volume 8, Number 1, Page 347
[8]
Li-juan Wang, Yan Zhang, and Chun-yang Zhang
Analytical Chemistry, 2013, Volume 85, Number 23, Page 11509
[9]
Jiří Fajkus
Clinica Chimica Acta, 2006, Volume 371, Number 1-2, Page 25
[10]
Ralph Hölzel, Nenad Gajovic-Eichelmann, and Frank F Bier
Biosensors and Bioelectronics, 2003, Volume 18, Number 5-6, Page 555
[11]
Yossi Weizmann, Fernando Patolsky, Eugenii Katz, and Itamar Willner
ChemBioChem, 2004, Volume 5, Number 7, Page 943
[12]
S. P. Rad’ko, S. A. Voronina, A. V. Gromov, O. V. Gnedenko, N. V. Bodoev, A. S. Ivanov, and K. N. Yarygin
Bulletin of Experimental Biology and Medicine, 2009, Volume 147, Number 6, Page 746
[13]
Jenny Steffen, Markus von Nickisch-Rosenegk, and Frank F. Bier
Lab on a Chip, 2005, Volume 5, Number 6, Page 665

Comments (0)

Please log in or register to comment.
Log in