DETECTING TARGET MOLECULES IN A SAMPLE

• US 20100197515A1
• Filed: 10/19/2007
• Published: 08/05/2010
• Est. Priority Date: 10/24/2006
• Status: Active Grant

• Current Assignee: Koninklijke Philips N.V.
• Original Assignee: Koninklijke Philips Electronics N.V. (Koninklijke Philips N.V.)
• Inventors: Stapert, Hendrik Roelof, Van Herpen, Maarten Marinus Johannes Wilhelm, Klunder, Derk Jan Wilfred
• Granted Patent: US 10,101,321 B2
• Time in Patent Office: Days
• US Class Current: 506/9
• CPC Class Codes: C40B 30/10 by measuring physical properties, e.g. mass
• CPC Class Codes: C40B 60/04 Integrated apparatus specially adapted for both screening libraries and identifying library members
• 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 33/5306 Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding

Abstract

The invention relates to detection the presence of a target molecule in a sample, wherein the sample is contacted with a substrate, the substrate subsequently being washed in a wash step. In particular, the invention relates to a method of detecting the presence of a target molecule in a sample, the method comprising: (a) contacting the sample (37) with a substrate having immobilized thereon probe molecules that specifically binds to the target molecule; (b) washing the substrate (38) in a wash step by a wash fluid in order to remove or dilute unbound target molecules; (c) detect the presence of resultant binding complexes (39) on the substrate to determine whether the target molecule is present in the sample. The wash fluid being substantially refractive index matched to the substrate.

Claims

1. Method of detecting the presence of a target molecule in a sample, the method comprising:

(a) contacting the sample (37) with a substrate having immobilized thereon probe molecules that specifically binds to the target molecule;

(b) washing the substrate (38) in a wash step by a wash fluid in order to remove or dilute unbound target molecules;

(c) detect the presence of resultant binding complexes (39) on the substrate to determine whether the target molecule is present in the sample;

wherein the wash fluid being substantially refractive index matched to the substrate.

2. The method according to claim 1, wherein the refractive index of the wash fluid is within the range of the refractive index of the substrate plus 10% to the refractive index of the substrate minus 10%.

3. The method according to claim 1, wherein the wash fluid is an aqueous solution of sugar in water, where the weight percentage of sugar is between 50% to 80%.

4. The method according to claim 1 further comprising at least one additional wash step prior to the wash step (b).

5. The method according to claim 1, wherein the substrate is a porous substrate.

6. The method according to claim 1, wherein the target molecules chemically bind to the probe molecules present in the substrate.

7. The method according to claim 1, wherein the presence of the target molecules is detected by detecting luminescence of the target molecules.

10. A kit for performing an assay, the kit comprising:

a wash fluid, the wash fluid being substantially index matched to a substrate;

instructions for using the wash fluid in a method according to claim 1.

8. Apparatus for preparing an assay, the apparatus comprising:

a handling unit (33) for receiving a substrate having immobilized thereon probe molecules that specifically binds to a target molecule and for contacting a sample with the substrate (3, 30-32);

a wash unit (35) for performing a wash step on the substrate using a wash fluid in order to remove or dilute unbound target molecules, the wash fluid being substantially refractive index matched to the substrate.

9. An optical detection system for detecting a target molecule in an assay, the detection system comprising:

a handling unit (33) for receiving a substrate having immobilized thereon probe molecules that specifically binds to a target molecule and for contacting a sample with the substrate;

a wash unit (34) for performing a wash step on the substrate using a wash fluid in order to remove or dilute unbound target molecules, the wash fluid being substantially refractive index matched to the substrate;

a detector (35, 36) for detecting the presence of resultant binding complexes on the substrate to determine whether the target molecule is present in the sample.