Methods and Compositions for Highly Sensitive Detection of Molecules

• US 20100329929A1
• Filed: 03/25/2010
• Published: 12/30/2010
• Est. Priority Date: 09/28/2004
• Status: Active Grant

• Current Assignee: Singulex, Inc.
• Original Assignee: Singulex, Inc.
• Inventors: Todd, John, Livingston, Richard A., Held, Douglas, Puskas, Robert, Goix, Philippe J.
• Granted Patent: US 8,685,711 B2
• Time in Patent Office: Days
• US Class Current: 422/69
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 2201/06113 Coherent sources; lasers
• CPC Class Codes: G01N 2201/062 LED's
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/68 involving proteins, peptides or amino acids involving lipoproteins G01N33/92

Abstract

Disclosed are methods, kits, and compositions for the highly sensitive detection of molecules. The methods, kits, and compositions are useful in determining concentrations of molecules in samples to levels of 1 femtomolar, 1 attomolar, or lower. The methods, kits, and compositions also allow the determination of concentration over a wide range, e.g., 7-log range, without need for sample dilution.

Claims

1. An analyzer system kit for detecting a single protein molecule in a sample, said kit comprising an analyzer and at least one label comprising a fluorescent moiety and a binding partner for said protein molecule, and wherein said analyzer comprises:

a) an electromagnetic radiation source for stimulating said fluorescent moiety;

b) a capillary flow cell for passing said label;

c) a source of motive force of moving said label in said capillary flow cell;

d) an interrogation space defined within said capillary flow cell for receiving electromagnetic radiation emitted from said electromagnetic source; and

e) an electromagnetic radiation detector operably connected to said interrogation space for measuring an electromagnetic characteristic of said stimulated fluorescent moiety;

wherein said fluorescent moiety is capable of emitting at least about 200 photons when simulated by a laser emitting light at the excitation wavelength of the moiety, wherein the laser is focused on a spot not less than about 5 microns in diameter that contains the moiety, and wherein the total energy directed at the spot by the laser is no more than about 3 microJoules.

2-49. -49. (canceled)