Red blood cell separation means for specific binding assays

• US 5,939,331 A
• Filed: 05/21/1993
• Issued: 08/17/1999
• Est. Priority Date: 03/10/1992
• Status: Expired due to Term

• Current Assignee: Quidel Corporation
• Original Assignee: Quidel Corporation
• Inventors: Burd, John, Rowley, Gerald, Pronovost, Allan, Miller, Steven
• Primary Examiner(s): Spiegel, Carol A.
• Application Number:
  US08/066,410
• Time in Patent Office: 2,279 Days
• Field of Search: 435/7.9, 435/7.92-7.95, 435/805, 435/962, 435/969, 435/970, 436/518, 436/527-535, 436/169-170, 436/810, 436/825, 422/56-58, 422/101
• US Class Current: 436/518
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/54388 based on lateral flow
• CPC Class Codes: G01N 33/556 Fixed or stabilised red blood cell
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: G01N 33/563 involving antibody fragments
• CPC Class Codes: G01N 33/80 involving blood groups or blood types or red blood cells white blood cells G01N33/56972
• CPC Class Codes: Y10S 435/962 Prevention or removal of interfering materials or reactants or other treatment to enhance results, e.g. determining or preventing nonspecific binding
• CPC Class Codes: Y10S 435/969 Multiple layering of reactants
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 436/825 Pretreatment for removal of interfering factors from sample

Abstract

A method and device for detecting the presence, absence or amount of an analyte in a whole blood sample is disclosed. The device comprises four zones, a sample receiving zone, a labeling zone, a capture zone and an absorbent zone. The sample receiving zone contains an irreversibly immobilized reagent that allows for removal of substantially all red blood cells from the whole blood sample. Flow through the device is via capillary migration and all of the dissolved or dispersed components in the sample flow at substantially equal rates and with relatively unimpaired flow through the device. The method involves the use of the device for detection of analyte in a whole blood sample.

Claims

1. A lateral flow device for determining of the presence, absence, or amount of an analyte in a whole blood sample, which device comprises:

i) a sample receiving zone comprising a first solid porous matrix with an irreversibly immobilized reagent that specifically binds red blood cell surface antigens and effects the removal of substantially all red blood cells (RBCs) from the whole blood sample to result in a substantially RBC-free fluid containing dissolved or dispersed components, said sample receiving zone being in lateral flow contact with

ii) a labeling zone comprising a second solid porous matrix with a mobilizable labeling component comprising a visible label coupled to a ligand selected from the group consisting of the analyte, a competitive analog of the analyte and a specific binder partner which specifically binds to the analyte and the analog, said labeling zone being in lateral flow contact with

iii) a capture zone comprising a third solid porous matrix with an immobilized capture reagent, said capture zone being in lateral flow contact with

iv) an absorbent zone;

wherein said sample receiving zone is separately prepared and positioned contiguous with the remainder of the device prior to application of the sample;

wherein the first, the second and third solid porous matrices are non-bibulous matrices; and

wherein all of said dissolved or dispersed components in said RBC-free fluid flow through the sample receiving zone, labeling zone and capture zone in a non-bibulous flow.

2. The device of claim 1 wherein the labeling component comprises the visible label coupled to the specific binding partner for said analyte.

3. The device of claim 1 wherein the labeling component comprises the visible label coupled to the competitive analog of the analyte.

4. The device of claim 1 wherein the first solid porous matrix is selected from the group consisting of paper, nitrocellulose, and nylon and wherein said matrix is converted to a non-bibulous material through the application of an effective concentration of methylated bovine serum albumin, and then the reagent that specifically binds red blood cell surface antigens is immobilized on said matrix, and said matrix is then lyophilized to provide for irreversible immobilization of the reagent.

5. The device of claim 1 wherein the reagent that specifically binds red blood cell surface antigens is selected from the group consisting of monoclonal antibody, polyclonal antibodies, and their F(ab'"'"')₂, Fab'"'"' and Fab fragments.

6. The device of claim 1 in which said first solid porous matrix, second solid porous matrix and third solid porous matrix each further comprise a non-woven fabric structure.

7. The device of claim 6 wherein said fabric has a spunlaced structure.

8. The device of claim 1 wherein said first solid porous matrix, second solid porous matrix and third solid porous matrix further each are comprised of a high density polyethylene sheet.

9. The device of claim 8, wherein said sheet has an open pore structure with a density, at 40% void volume, of 0.57 gm/cc and an average pore diameter of 1 to 250 micrometers.

10. The device of claim 1 wherein said zones have a pore size of about 1-100 micrometers.

11. A method to determine the presence, absence, or amount of analyte in a whole blood sample which method comprises the steps of:

(a) contacting the whole blood sample with the device of claim 1; and

(b) visually observing the binding of the labeling component in the third solid porous matrix as a function of the presence or absence or amount of the analyte in the whole blood sample.