Integrated high throughput system for the analysis of biomolecules

• US 8,372,279 B2
• Filed: 12/08/2011
• Issued: 02/12/2013
• Est. Priority Date: 01/18/2001
• Status: Expired due to Term

• Current Assignee: Intrinsic Bioprobes, Inc. (Perspecta, Inc.)
• Original Assignee: Intrinsic Bioprobes, Inc. (Perspecta, Inc.)
• Inventors: Tubbs, Kemmons, Gruber, Karl F., Nelson, Randall W.
• Primary Examiner(s): Therkorn, Ernest G
• Application Number:
  US13/315,163
• Publication Number:
  US 20120085692A1
• Time in Patent Office: 432 Days
• Field of Search: 210/635, 210/656, 210/659, 210/198.2, 210/502.1, 961/01, 422/70, 422/100, 422/101
• US Class Current: 210/198.2
• CPC Class Codes: B01D 15/3804 Affinity chromatography
• CPC Class Codes: B01J 20/28097 being coated, filled or plugged with specific compounds
• CPC Class Codes: B01J 20/281 Sorbents specially adapted for preparative, analytical or investigative chromatography
• CPC Class Codes: B01J 20/286 Phases chemically bonded to a substrate, e.g. to silica or to polymers
• CPC Class Codes: B01J 20/32 Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
• CPC Class Codes: B01J 20/3204 Inorganic carriers, supports or substrates
• CPC Class Codes: B01J 20/3208 Polymeric carriers, supports or substrates
• CPC Class Codes: B01J 20/3242 Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
• CPC Class Codes: B01J 20/3259 comprising at least two different types of heteroatoms selected from nitrogen, oxygen or sulfur with at least one silicon atom
• CPC Class Codes: B01J 20/3265 with an organic functional group containing a metal, e.g. a metal affinity ligand
• CPC Class Codes: B01J 20/3272 Polymers obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3274 Proteins, nucleic acids, polysaccharides, antibodies or antigens
• CPC Class Codes: B01J 20/3282 Crosslinked polymers
• CPC Class Codes: B01J 2220/54 Sorbents specially adapted for analytical or investigative chromatography
• CPC Class Codes: B01J 2220/64 In a syringe, pipette, e.g. tip or in a tube, e.g. test-tube or u-shape tube in columns B01J2220/58
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: G01N 2570/00 Omics, e.g. proteomics, glycomics or lipidomics; Methods of analysis focusing on the entire complement of classes of biological molecules or subsets thereof, i.e. focusing on proteomes, glycomes or lipidomes
• CPC Class Codes: G01N 30/02 Column chromatography
• CPC Class Codes: G01N 30/6095 Micromachined or nanomachined, e.g. micro- or nanosize
• 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 33/6803 : General methods of protein analysis not limited to specific proteins or families of proteins
• CPC Class Codes: G01N 33/6845 : Methods of identifying protein-protein interactions in protein mixtures
• CPC Class Codes: H01J 49/164 : Laser desorption/ionisation, e.g. matrix-assisted laser desorption/ionisation [MALDI] sample holders H01J49/0418
• CPC Class Codes: H01J 49/40 : Time-of-flight spectrometers H01J49/36 takes precedence
• CPC Class Codes: Y10T 436/255 : including use of a solid sorbent, semipermeable membrane, or liquid extraction

Abstract

Described is an affinity microcolumn comprising a high surface area material, which has high flow properties and a low dead volume, contained within a housing and having affinity reagents bound to the surface of the high surface area material that are either activated or activatable. The affinity reagents bound to the surface of the affinity microcolumn further comprise affinity receptors for the integration into high throughput analysis of biomolecules.

Claims

1. An affinity microcolumn comprising a porous molecular trap having a high surface area material comprising silica, which has high flow properties and a low dead volume, contained within a housing and affinity reagents bound to the surface of the high surface area material, wherein the affinity reagents are either activated or activatable and the high surface area is formed by at least one of chemical etching and electro etching.

2. The affinity microcolumn of claim 1 wherein the porous molecular trap is formed by molding.

3. The affinity microcolumn of claim 1 wherein the affinity reagents that are bound to the surface of the high surface area material further comprise affinity receptors bound to the affinity reagents.

4. The affinity microcolumn of claim 3 further comprising a tethering molecule that is activated or activatible and binds the affinity receptors to the affinity reagents.

5. The affinity microcolumn of claim 3 further comprising an amplification media that is activated or activatible and is interposed between the affinity reagents and the affinity receptors, where the amplification media allows a high density of affinity receptors to be bound to the affinity reagents than in the absence of the amplification media.

6. The affinity microcolumn of claim 5 wherein the amplification media comprises at least one of a biological polymer, a non-biological organic polymer, and an inorganic polymer.

7. The affinity microcolumn of claim 3 further comprising an amplification media interposed between the affinity reagents and the affinity receptors, where the amplification media allows better access by an analyte to the affinity receptors than in the absence of the amplification media.

8. The affinity microcolumn of claim 7 wherein the amplification media comprises at least one of a biological polymer, a non-biological organic polymer, and an inorganic polymer.

9. The affinity microcolumn of claim 1 wherein the housing is a micropipette.

10. The affinity microcolumn of claim 1 wherein the housing is a manifold having more than one microchannel, the porous molecular trap fitting into at least one of the microchannels.

11. The affinity microcolumn of claim 9 wherein the housing further comprises at least one projection that supports the porous molecular trap.

12. The affinity microcolumn of claim 9 wherein the housing further comprises at least one projection in at least one of the microchannels, the projection supporting the porous molecular trap.