Fluorescence polarization assay

a fluorescence polarization and assay technology, applied in the field of fluorescence polarization assay, can solve the problems of limited antibody-based molecular recognition molecules, and therefore useful in diagnostic assays, and the platform that has typically been used to identify and quantify aptamers bound to analytes is very limited

a fluorescence polarization and assay technology, applied in the field of fluorescence polarization assay, can solve the problems of limited antibody-based molecular recognition molecules, and therefore useful in diagnostic assays, and the platform that has typically been used to identify and quantify aptamers bound to analytes is very limited

US20070054308A1Inactive Publication Date: 2007-03-08APPL BIOSYSTEMS INC
35 Cites 0 Cited by

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Fluorescence polarization assay
  • Fluorescence polarization assay
  • Fluorescence polarization assay

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0091] A. Methods for assessing binding of proteins to aptamer-coupled beads

[0092] The polarization anisotropy of dye-labeled aptmers changes with time upon binding to a specific protein. Changes in fluorescence anisotropy of 3 aptamers bound to a solid support upon binding of specific proteins was measured.

[0093] The three fluorophore labeled aptamers used in these experiments were anti-thrombin aptamer (FAM-labeled, a 15 mer), which is specific for thrombin from human plasma, 518-aptamer (FAM-labeled, a 60 mer), which is specific for GP120MN, an HIV-1 protein and 650-aptamer (FAM-labeled, a 60 mer), which is specific for recombinant human FGF basic protein.

[0094] 1. Anti-Thrombin Aptamer for Thrombin

[0095] Binding of thrombin to FAM-labeled anti-thrombin aptamers was measured. 5 micron silica particles were purchased from Bangs Laboratories (Ohio, USA) and pretreated with 1N NaCI solution at 100° C. for 1 hour. The silica particles were then treated for 30 minutes with 3-amino...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
diameteraaaaaaaaaa
diameteraaaaaaaaaa
diameteraaaaaaaaaa
Login to View More

Abstract

The present invention relates to methods for detecting the presence of one or more analytes of interest in a sample by measuring changes in fluorescence anisotropy as a result of binding of the analytes to specific aptamers. The aptamers are immobilized on a solid support and may be in the form of an array.

Description

REFERENCE TO RELATED APPLICATIONS [0001] The present application is a continuation of U.S. application Ser. No. 10 / 628,879, filed Jul. 28, 2003, which claims priority under 35 U.S.C. §119(e) to U.S. Provisional Application No. 60 / 401,021, filed Aug. 2, 2002, the entireties of both of which are incorporated by reference.FIELD OF THE INVENTION [0002] The present invention relates to methods for detecting the binding of analytes of interest to aptamers by measuring changes in fluorescence anisotropy. DESCRIPTION OF THE RELATED ART [0003] The introduction of immunoassays in 1959 and the enzyme-linked immunosorbent assay (ELISA) in 1971 revolutionized clinical diagnostic medicine. Until recently, molecules capable of molecular recognition, and therefore useful in diagnostic assays, have been limited to antibodies. The recent development of in vitro methods to select high affinity ligands by combinatorial chemistry methodologies promises unique novel therapeutics and diagnostic reagents. ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
08 Mar 2007
Publication
US20070054308A1
IPC
C12Q1/68; G01N33/543; G01N33/53; C12M1/00; G01N21/76; G01N21/78; G01N33/552; G01N33/58; G01N37/00
CPC
C12Q1/6837; G01N33/54313; G01N33/582; C12Q2561/119; C12Q2525/205
Inventors
SEKAR, MICHAEL M. A.; GREENFIELD, I. LAWRENCE