TOWARDS A NEW HOMOGENEOUS IMMUNOASSAY FOR GONADOTROPIN-RELEASING HORMONE BASED ON TIME-RESOLVED FLUORESCENCE ANISOTROPY
Peter D. Dowd, Jan Karolin, Carol Trager-Cowan, David J. S. Birch, William H. Stimson
2011
Abstract
A new method for detecting gonadotropin-releasing hormone type 1 (GnRH-1) is proposed. The method is based on a fluorescence polarisation immunoassay where the GnRH-1 vies for the binding site on the anti-GnRH (antibody 7B10.1D10) with a labelled peptide consisting of a fragment of GnRH labelled with fluorescein. Time-resolved anisotropy decays indicate increased correlation times for the labelled fragment in the presence of 7B10.1D10 compared with the correlation times for the labelled peptide fragment alone. When GnRH-1 is added to the solution of the labelled peptide in the presence of 7B10.1D10 it has the effect of reducing the correlation times. The results demonstrate the possibility of introducing the labelled fragment as a competitor to the antigen in the fluorescence polarisation immunoassay for GnRH-1 and, more generally, in the case where there is a disadvantage in labelling the antigen, such as GnRH-1, with a fluorescent probe.
References
- Apperson, K., Karolin, J., Martin, R. W., Birch, D. J. S., 2009 Nanoparticle Metrology Standards Based on Time-resolved Fluorescence Anisotropy of Silica Colloids. Meas. Sci. Technol. 20, 025310
- Chen, A., Kaganovsky, E., Rahimipour, S., Ben-Aroya, N., Okon, E., Koch, Y., 2002 Two Forms of Gonadotropin-releasing Hormone (GnRH) are Expressed in Human Breast Tissue and Over expressed in Breast Cancer: A Putative Mechanism for the Antiproliferative Effect of GnRH by DownRegulation of Acidic Ribosomal Phosphoproteins P1 and P21. Cancer Research. 62, 1036-1044
- Conn, P. M., Crowley, W. F., Jr. 1991 Gonadotropin releasing Hormone and its Analogues. N. Engl. J. Med. 324, 93-103
- Khan M. A. H., Ferro V. A., Stimson W. H. 2003 Use of a Highly Specific Monoclonal Antibody Against the Central Variable Amino Acid Sequence of Mammalian Gonadotropin Releasing Hormone to Evaluate GnRH-1 Tissue Distribution Compared with GnRH-1 Binding Sites in Adult Male Rats. AJRI. 49, 239-248
- Lakowicz J. R., 2006 Principles of Fluorescence Spectroscopy, 3rd ed. Kluwer Academic/Plenum Publishers, New York City, New York
- Price, C. P., 2001 Clinical Review: Point of Care Testing. BMJ, 322, 1285-1288
- Spencer, R. D., Toledo, F.B., Williams, B.T., and Yoss, N.L 1973 Design, Construction, and 2 Applications for an Automated Flow-cell Polarization Fluorometer with Digital Read Out-Enzyme-Inhibitor (Antitrypsin) Assay and Antigen-antibody (Insulin-insulin Aantiserum) Assay. Clin. Chem. 19, 838-844
- Tanriverdi, F., Silveira, L., Ganzalez-Martinez, D., Hu,Y., Bouloux, P., Kelestimur, F. 2004 Gonadotropinreleasing Hormone Type-I (GnRH-1) Expression in Peripheral Lymphocytes Aand Possible Immune Action. Erciyes Tip Dergisi (Erciyes Medical Journal). 26 (3), 105-112
- Terpetsching, E., Szmacinski, H., Lakowicz J. R., 1995 Fluorescence Polarization Immunoassay of a HighMolecular-Weight Antigen Based on a Long-Lifetime Ru-Ligand Complex. Anal. Biochem. 227, 140-147
Paper Citation
in Harvard Style
Dowd P., Karolin J., Trager-Cowan C., S. Birch D. and Stimson W. (2011). TOWARDS A NEW HOMOGENEOUS IMMUNOASSAY FOR GONADOTROPIN-RELEASING HORMONE BASED ON TIME-RESOLVED FLUORESCENCE ANISOTROPY . In Proceedings of the International Conference on Biomedical Electronics and Devices - Volume 1: BIODEVICES, (BIOSTEC 2011) ISBN 978-989-8425-37-9, pages 184-188. DOI: 10.5220/0003152001840188
in Bibtex Style
@conference{biodevices11,
author={Peter D. Dowd and Jan Karolin and Carol Trager-Cowan and David J. S. Birch and William H. Stimson},
title={TOWARDS A NEW HOMOGENEOUS IMMUNOASSAY FOR GONADOTROPIN-RELEASING HORMONE BASED ON TIME-RESOLVED FLUORESCENCE ANISOTROPY},
booktitle={Proceedings of the International Conference on Biomedical Electronics and Devices - Volume 1: BIODEVICES, (BIOSTEC 2011)},
year={2011},
pages={184-188},
publisher={SciTePress},
organization={INSTICC},
doi={10.5220/0003152001840188},
isbn={978-989-8425-37-9},
}
in EndNote Style
TY - CONF
JO - Proceedings of the International Conference on Biomedical Electronics and Devices - Volume 1: BIODEVICES, (BIOSTEC 2011)
TI - TOWARDS A NEW HOMOGENEOUS IMMUNOASSAY FOR GONADOTROPIN-RELEASING HORMONE BASED ON TIME-RESOLVED FLUORESCENCE ANISOTROPY
SN - 978-989-8425-37-9
AU - Dowd P.
AU - Karolin J.
AU - Trager-Cowan C.
AU - S. Birch D.
AU - Stimson W.
PY - 2011
SP - 184
EP - 188
DO - 10.5220/0003152001840188