Colloid compositions for solid phase biomolecular analytical, preparative and identification systems

• US 6,921,637 B2
• Filed: 05/04/2001
• Issued: 07/26/2005
• Est. Priority Date: 05/04/2000
• Status: Expired due to Fees

• Current Assignee: Center For Blood Research Incorporated
• Original Assignee: CBR Institute for Biomedical Research Inc.
• Inventors: McCormick, William, Fici, Dolores A., Audeh, Zuheir L.
• Primary Examiner(s): Chin, Chris
• Assistant Examiner(s): Padmanabhan, Kartic
• Application Number:
  US09/848,777
• Publication Number:
  US 20020015958A1
• Time in Patent Office: 1,544 Days
• Field of Search: 435/4, 435/6, 435/7.1, 435/7.9, 435/7.92, 435/287.1, 436/164, 436/172, 436/518, 436/524, 436/532, 436/536, 422/68.1
• US Class Current: 506/39
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00466 in a slurry
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00545 Colours
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00641 the porous medium being continuous, e.g. porous oxide substrates
• CPC Class Codes: B01J 2219/00648 by the use of solid beads
• CPC Class Codes: B01J 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01L 3/5023 with a sample being transported to, and subsequently stored in an absorbent for analysis
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: G01N 33/54346 Nanoparticles
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing

Abstract

A liquid composition comprising a colloidal suspension of a biomolecule-binding matrix material (preferably nitrocellulose) dispersed in a liquid, with particles of the matrix material being of a defined particle size, and replicate copies of a biomolecule, e.g., protein or nucleic acid probes, which are distributed, preferably uniformly, throughout the colloidal suspension and are bound to the matrix material particles, is disclosed. The liquid composition of the invention can be used directly for sample analysis or preparation of biomolecules, or aliquots of the composition can be spotted onto a support to form a microporous matrix system or microarray for analysis or preparation of biomolecules. Compositions and microarrays according to the invention are useful in any type of analytical or preparative procedure relating to biomolecules. They are particularly useful, e.g., in methods for detecting a biomolecule analyte in a liquid sample, methods for determining the presence of a particular nucleic acid sequence within a liquid sample and methods for determining the presence of a drug candidate molecule in a liquid sample. The invention further comprises kits for practicing the various methods of the invention.

Claims

1. A liquid composition comprising

a colloidal suspension of a biomolecule-binding matrix material uniformly dispersed in a liquid, wherein colloidal particles of said matrix material in said colloidal suspension are of a defined particle size of less than about 1 μ

m in diameter, and wherein said biomolecule-binding matrix material is selected from the group consisting of nitrocellulose, polyvinyl difluoride and activated nylon;

replicate copies of a biologically active labeled biomolecule, wherein said biomolecules are distributed throughout said colloidal suspension and are bound to said matrix material particles and wherein said biomolecule is selected from the group consisting of a protein, peptide and oligopeptide;

one or more reference dyes distributed throughout said colloidal suspension; and

a blocking biomolecule, wherein said blocking biomolecule blocks sites on said biomolecule-binding matrix material not occupied by said biomolecule.

2. The liquid composition of claim 1, wherein the reference dye is a marker for determining the quantity of biomolecule deposited in a spot on a microarray.

3. The liquid composition of claim 1, wherein said particles of matrix material have a diameter of less than 0.5 μ

m.

4. The liquid composition of claim 1, wherein said particles of matrix material have a diameter of less than 0.25 μ

m.

5. The liquid composition of claim 1 wherein the biomolecule is a protein.

6. The liquid composition of claim 5 wherein the biomolecule-binding matrix material is nitrocellulose.

7. The liquid composition of claim 1, wherein more than one species of biomolecule is distributed throughout said colloidal suspension and bound to said matrix material particles.

8. The liquid composition of claim 7, wherein said more than one species of biomolecule comprise two or more different biomolecule probes.

9. The liquid composition of claim 1, wherein said binding of said biomolecules is covalent binding.

10. The liquid composition of claim 1, wherein said binding of said biomolecules is non-covalent binding.

11. The liquid composition of claim 1, wherein said binding of said biomolecules is electrostatic binding.

12. The liquid composition of claim 1, wherein said binding of said biomolecules is adsorption onto a surface of said matrix material particles.

13. The liquid composition of claim 1, wherein the reference dye is used to track the concentration of matrix material during the preparation of the liquid composition.