Nanoporous membrane immunosensor

• US 7,341,696 B2
• Filed: 10/05/2006
• Issued: 03/11/2008
• Est. Priority Date: 07/12/2000
• Status: Expired due to Fees

• Current Assignee: Navy The USA As Represented By The Secretary of The
• Original Assignee: the united states of america as represented by the secretary of the navy
• Inventors: Lee, Gil U, Yanavich, Carolyn
• Primary Examiner(s): Le; Long V.
• Assistant Examiner(s): Counts; Gary W.
• Application Number:
  US11/538,987
• Publication Number:
  US 20070264706A1
• Time in Patent Office: 523 Days
• Field of Search: 422/57, 422/58, 422/68.1, 422/100, 422/101, 422/186.1, 435/6, 435/7.1, 435/287.1, 435/287.2, 435/973, 436/518, 436/164, 210/348, 210/455, 210/600
• US Class Current: 422/534
• CPC Class Codes: G01N 33/537 with separation of immune complex from unbound antigen or antibody
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding

Abstract

A sensor for a selected analyte in a test sample has (a) a semipermeable membrane with pores for retaining the analyte, where the membrane has been chemically modified by attachment of membrane modifiers; (b) immunoassay labels which have label binding ligands where these label binding ligands will have a binding affinity for the membrane modifiers in the presence of the analyte, and a measurably different binding affinity for the membrane modifiers in the absence of the analyte; and (c) a label detecting system, for detecting the presence of the labels on the membrane.

Claims

1. A sensor for independently detecting a plurality of analytes in a test solution, comprising:

a test vessel;

a semipermeable membrane with pores for retaining the analyte, dividing said test vessel into a first volume and a second volume, wherein a surface of said membrane is chemically modified by attachment of at least two distinct membrane surface modifiers on at least a side facing said first volume but not within said pores, wherein said at least two distinct membrane surface modifiers are patterned into an array of distinct regions on said membrane surface;

at least two distinct groups of immunoassay labels disposed within said first volume, wherein each of said groups of immunoassay labels has distinct label binding ligands where these label binding ligands will have a binding affinity for one of said distinct membrane surface modifiers in the presence of the analyte, and a measurably different binding affinity for said one of said distinct membrane surface modifiers in the absence of the analyte;

a pressure source, for driving said test solution from said first volume into said second volume; and

a label detecting system, for detecting the presence or absence of said labels in each of said regions on said membrane.

2. A sensor for independently detecting a plurality of analytes in a test solution, comprising:

a test vessel;

a semipermeable membrane with pores for retaining the analyte, dividing said test vessel into a first volume and a second volume, said pores are selected to prevent the analyte from passing into or through said semipermeable membrane;

wherein said membrane is chemically modified by attachment of at least two distinct membrane modifiers on at least a side facing said first volume but not within said pores;

wherein said at least two distinct membrane modifiers are patterned into an array of distinct regions on said membrane;

at least two distinct groups of assay labels disposed within said first volume, wherein each of said groups of assay labels has distinct label modifiers, said label modifiers having a binding affinity for one of said distinct membrane modifiers in the presence of the analyte, and a measurably different binding affinity for said one of said distinct membrane modifiers in the absence of the analyte;

a pressure source, for driving said test solution from said first volume into said second volume; and

a label detecting system, for detecting the presence or absence of said labels in each of said regions on said membrane.

3. The sensor of claim 2, wherein said membrane supports a 100 kPa pressure load and said membrane is functionalized with a binder at the surface of said membrane in order for said membrane to act as a sensor.

4. The sensor of claim 2, wherein said membrane has pores not greater than 10 nm in diameter.

5. The sensor of claim 2, wherein said membrane has a pore density of at least 10¹⁵/m².

6. The sensor of claim 2, wherein said membrane is essentially flat and optically translucent when wet.

7. The sensor of claim 6, wherein said membrane remains translucent, and the shape of said membrane remains flat even under pressure associated with flow of solution through said membrane.

8. The sensor of claim 2, wherein said membrane is an aluminum oxide membrane.

9. The sensor of claim 2, wherein an active surface of said membrane is modified with a biotin-polyethylene-glycol (PEG) using a polyethyleneimine (PEI) layer.

10. The sensor of claim 2, wherein said pores allow a solvent to pass through while preventing flow of said binder, analyte, or assay labels.

11. The sensor of claim 2, wherein said membrane has pores not greater than 20 nm in diameter.