Method of performing assays for biomolecules and solid supports for use in such methods

• US 5,424,219 A
• Filed: 10/25/1991
• Issued: 06/13/1995
• Est. Priority Date: 10/25/1991
• Status: Expired due to Fees

• Current Assignee: Cytech Biomedical Incorporated
• Original Assignee: Cytech Biomedical Incorporated
• Inventors: Jirikowski, Gustav F.
• Primary Examiner(s): Scheiner, Toni R.
• Assistant Examiner(s): Chin, Christopher L.
• Application Number:
  US07/782,934
• Time in Patent Office: 1,327 Days
• Field of Search: 528/491, 528/494, 528/495, 528/497, 528/498, 528/502, 156/654, 156/668, 435/4, 435/6, 436/518, 436/531, 436/800, 436/805, 436/809, 436/807, 522/1
• US Class Current: 436/518
• CPC Class Codes: G01N 33/5436 with ligand physically entrapped within the solid phase liposomes G01N33/5432; immunological test elements G01N33/54386
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/545 Synthetic resin
• CPC Class Codes: G01N 33/56988 HIV or HTLV
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays

Abstract

Methods of performing an assay for biomolecules and solid supports for use in such assays are disclosed. An activated polymeric material is used as a solid support for binding biomolecules. In a preferred form, the support is made of a generally hydrophobic, nonporous polymer which has been activated by treatment with solvents or by mechanical means to enhance the binding characteristics of the support. The support can be made of a copolymer, such as a copolymer of polystyrene and polybutadiene. Detection of bound biomolecules is preferably performed by means of staining reactions and gray level scanning. Emulsifying agents, such as detergents, can be applied to the surface of the support to substantially inhibit additional materials from being bound to the support.

Claims

1. A method of performing an assay for biomolecules in a sample comprising:

providing a solid support of polymeric composition consisting essentially of at least two suitable polymeric components selected from the group consisting of polymers and copolymers, said suitable polymeric components capable of dissolving at different rates when exposed to a solvent composition, so that one of said suitable polymeric components dissolves more quickly and the other of said suitable polymeric components dissolves less readily when said solid support is exposed to said solvent composition,

exposing said solid support to said suitable solvent composition in a region of said solid support to remove a portion of the polymeric components in said region with said solvent composition to increase the exposed surface area of the region,

contacting the sample to said solid support and allowing binding region of the solid support,

removing any excess sample from the solid support,

contacting the solid support with a detectable reagent that specifically binds to the biomolecules,

removing excess detectable reagent from the solid support, and

detecting the detectable reagent on the solid support to determine the presence of biomolecules bound to said region of the solid support.

2. The method of claim 1 in which the solid support has been activated by treating said region with an organic solvent to partially dissolve the polymeric material.

3. The method of claim 2 in which the solvent is selected from the group consisting of ethers, alcohols, ketones, aldehydes, acids, bases, olefins, linear and cyclic hydrocarbons, and halogenated hydrocarbons.

4. The method of claim 1 in which the polymeric composition comprises a member selected from the group consisting of polystyrene and polyvinylchloride.

5. The method of claim 1 in which the polymeric composition is a copolymer of styrene and butadiene.

6. The method of claim 1 in which the activated region is opaque.

7. The method of claim 1 in which the biomolecule is detected by using a staining dye to indicate the presence of the biomolecule and using gray level scanning to measure the intensity of staining.

8. The method of claim 1 in which the biomolecule is detected by using a substantially water-insoluble staining dye.

10. The method of claim 1 in which the biomolecule is covalently bound to the surface of the activated region of the support after binding the biomolecule.

11. The method of claim 10 in which the biomolecule is embedded in the activated region of the support by exposing the activated region of the support binding the biomolecule to ultraviolet light.

12. The method of claim 1 in which the support is formed in the shape of a substantially flat plate.

13. A method of performing an assay for biomolecules as in claim 1 comprising:

subsequently applying an emulsifying agent to the surface of the solid support after the biomolecule is bound to the solid support to substantially inhibit additional materials from being bound to the support.

14. The method of claim 13 in which the polymeric material comprises a member selected from the group consisting of polypropylene, polystyrene, polyvinylchloride, polyamide, polycarbonate, epoxides, methacrylates, and melamine resin.

15. The method of claim 13 in which the polymeric material is a copolymer.

16. The method of claim 15 in which the polymeric material is a copolymer of styrene and butadiene.

17. The method of claim 13 in which the emulsifying agent is a detergent.

18. The method of claim 13 in which the biomolecule is detected by using a staining dye to indicate the presence of the biomolecule and using gray level scanning to measure the intensity of staining.

9. The method 1 in which the biomolecule is a member selected from the group consisting of proteins, nucleic acids, amino acids, peptides, amines, and nucleotides.

19. The method 13 in which the biomolecule is a member selected from the group consisting of proteins, nucleic acids, amino acids, peptides, amines, and nucleotides.