Device and method of seperating and assaying whole blood

• US 5,558,834 A
• Filed: 05/31/1995
• Issued: 09/24/1996
• Est. Priority Date: 10/03/1991
• Status: Expired due to Term

• Current Assignee: Bayer Corporation (Bayer AG)
• Original Assignee: Bayer Corporation (Bayer AG)
• Inventors: Chu, Amy H., Stover, Lon R.
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): Pyon, Harold Y.
• Application Number:
  US08/454,614
• Time in Patent Office: 482 Days
• Field of Search: 422/55-58, 422/61, 422/68.1, 422/69, 422/73, 435/7.1, 435/13, 435/14, 436/5, 436/16, 436/45, 436/63, 436/66, 436/169, 436/177, 436/178, 436/526, 436/531, 436/501, 436/518, 436/524, 436/810, 210/483, 210/488, 210/496, 210/504, 210/506, 427/2.13, 427/244
• US Class Current: 422/422
• CPC Class Codes: G01N 33/525 Multi-layer analytical elements
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick

Abstract

An improved device and method of separating the cellular components of whole blood from plasma or serum and assaying the plasma or serum for a predetermined soluble constituent are disclosed. The device includes a filter pad, that separates the cellular components of whole blood from the serum or plasma, and a test pad, that assays the serum or plasma for a predetermined soluble constituent. The filter pad effectively separates and retains cellular components of the whole blood sample, thereby eliminating assay interference by the cellular components of whole blood. The method includes contacting the whole blood with a test device including a filter pad comprising a suitable carrier matrix homogeneously incorporating therein a separating reagent composition comprising a) separating reagent, like an agglutinin, such as a blood type nonspecific lectin; a coagulant, such as a thrombin or a thrombin-like compound; or a mixture thereof, and b) a nonhemolytic surfactant, like an ethoxylated or propoxylated nonionic or anionic surfactant, such that the cellular components of the whole blood are separated from the plasma or serum as the blood permeates through the filter pad, The essentially cell-free plasma or serum then saturates a test pad that is in contact with the filter pad. After the plasma or serum saturates the test pad, the test pad is examined for a qualitative or quantitative response to a predetermined soluble constituent of the whole blood.

Claims

1. A device for separating undiluted plasma or serum from an undiluted whole blood sample including a filter pad consisting essentially of a single carrier matrix having homogeneously incorporated therein from about 5 to about 50 units of a blood cell agglutinizing agent per cm³ of the carrier matrix, or from about 40 to about 100 NIH units of a blood cell coagulating agent per cm³ of the carrier matrix, or a mixture thereof, and from about 0.3% to about 1.5% by weight of the carrier matrix of a nonhemolytic surfactant, to agglutinate or coagulate the cellular components from the undiluted whole blood sample in the area of the carrier matrix while allowing a quantity of the undiluted plasma or serum portion of the undiluted whole blood sample to chromatographically flow completely through the carrier matrix separate from the cellular components of the blood, wherein the agglutinizing agent is a lectin, the coagulating agent is a thrombin compound and the nonhemolytic surfactant is selected from the group consisting of octoxynol 40, PEG-36 castor oil, C_11-15 pareth-12, C_12-15 pareth-9, ceteareth-27, ceteth-30, laureth-23, oleth-15, PEG-20 laurate, PEG-36 oleate, PEG-44 sorbitan laurate, PEG-30 stearate, steareth-27, PEG-11 cocamide, PEG-15 cocamine, PEG-15 tallow amine, PEG-20 hydrogenated castor oil, PPG-5-ceteth 20, PPG-myreth-11, steareth-10, trideceth-15, C_11-15 pareth-7 carboxylic acid, C_12-13 pareth-5 carboxylic acid, C_12-15 pareth-7 carboxylic acid, ceteareth-25 carboxylic acid, deceth-7 carboxylic acid, isosteareth-6 carboxylic acid, isosteareth-11 carboxylic acid, coceth-7 carboxylic acid, laureth-5 carboxylic acid, laureth-10 carboxylic acid, trideceth-4 carboxylic acid, trideceth-7 carboxylic acid, trideceth-15 carboxylic acid, trideceth-19 carboxylic acid, C_12-15 pareth-2 phosphate, ceteareth-4 phosphate, deceth-4 phosphate, dilaureth-10 phosphate, dioleth-8 phosphate, laneth-4 phosphate, laureth-3 phosphate, laureth-8 phosphate, nonoxynol-6 phosphate, nonoxynol-9 phosphate, nonoxynol-10 phosphate, nonyl nonoxynol-7 phosphate, nonyl nonoxynol-9 phosphate, nonyl nonoxynol-10 phosphate, nonyl nonoxynol-15 phosphate, nonyl nonoxynol-24 phosphate, oleth-3 phosphate, oleth-4 phosphate, oleth-10 phosphate, oleth-20 phosphate, potassium octoxynol-12 phosphate, PPG-5-ceteth phosphate, PPG-10 cetyl ether phosphate, triceteareth-4 phosphate, triceteth-5 phosphate, trideceth-6 phosphate, trilaneth-4 phosphate, trilaureth-4 phosphate, trioleth-8 phosphate, and combinations thereof,

wherein the amount of agglutinin or coagulant incorporated in the carrier matrix is about two to about five times less than the amount of agglutinin or coagulant used in the absence of a nonhemolytic surfactant.

2. An analyte test device that separates cellular and particulate components from a liquid test sample and that determines the presence or concentration of a soluble component of the liquid test sample consisting essentially of:

a support strip;

a test pad secured to the support strip, the test pad comprising a substrate material incorporating an indicator reagent composition that undergoes a detectable change in response to the soluble component of the liquid test sample when in contact with the liquid test sample; and

a filter pad of sufficient size to separate the cellular and particulate components from the liquid test sample, the filter pad consisting essentially of a single carrier matrix having homogeneously incorporated therein from about 5 to about 50 units of an agglutinin per cm³ of the carrier matrix or from about 40 to about 100 NIH units of a coagulant per cm³ of the carrier matrix, or a mixture thereof, and from about 0.3% to about 1.5% by weight of the carrier matrix of a nonhemolytic surfactant selected from the group consisting of octoxynol 40, PEG-36 castor oil, C_11-15 pareth-12, C_12-15 pareth-9, ceteareth-27, ceteth-30, laureth-23, oleth-15, PEG-20 laurate, PEG-36 oleate, PEG-44 sorbitan laurate, PEG-30 stearate, steareth-27, PEG-11 cocamide, PEG-15 cocamine, PEG-15 tallow amine, PEG-20 hydrogenated castor oil, PPG-5-ceteth 20, PPG-myreth-11, steareth-10, trideceth-15, C_11-15 pareth-7 carboxylic acid, C_12-13 pareth-5 carboxylic acid, C_12-15 pareth-7 carboxylic acid, ceteareth-25 carboxylic acid, deceth-7 carboxylic acid, isosteareth-6 carboxylic acid, isosteareth-11 carboxylic acid, coceth-7 carboxylic acid, laureth-5 carboxylic acid, laureth-10 carboxylic acid, trideceth-4 carboxylic acid, trideceth-7 carboxylic acid, trideceth-15 carboxylic acid, trideceth-19 carboxylic acid, C_12-15 pareth-2 phosphate, ceteareth-4 phosphate, deceth-4 phosphate, dilaureth-10 phosphate, dioleth-8 phosphate, laneth-4 phosphate, laureth-3 phosphate, laureth-8 phosphate, nonoxynol-6 phosphate, nonoxynol-9 phosphate, nonoxynol-10 phosphate, nonyl nonoxynol-7 phosphate, nonyl nonoxynol-9 phosphate, nonyl nonoxynol-10 phosphate, nonyl nonoxynol-15 phosphate, nonyl nonoxynol-24 phosphate, oleth-3 phosphate, oleth-4 phosphate, oleth-10 phosphate, oleth-20 phosphate, potassium octoxynol-12 phosphate, PPG-5-ceteth phosphate, PPG-10 cetyl ether phosphate, triceteareth-4 phosphate, triceteth-5 phosphate, trideceth-6 phosphate, trilaneth-4 phosphate, trilaureth-4 phosphate, trioleth-8 phosphate, and combinations thereof;

wherein the filter pad is positioned such that the liquid test sample first contacts the filter pad to chromatographically flow through the filter pad to contact the test pad, and

wherein the amount of agglutinin or coagulant incorporated in the carrier matrix is about two to about five times less than the amount of agglutinin or coagulant used in the absence of a nonhemolytic surfactant.

3. The analyte test device of claim 2 wherein the carrier matrix is filter paper;

and wherein the agglutinin is a lectin from Solanum tuberosum, the coagulant is bovine thrombin and the nonhemolytic surfactant is selected from the group consisting of octoxynol 40, PEG-36 castor oil, PPG-5-ceteth-10 phosphate, and combinations thereof.

4. The analyte test device of claim 2 wherein the liquid test sample is a biological sample.

5. The analyte test device of claim 4 wherein the biological sample is whole blood.

6. The analyte test device of claim 5 wherein the indicator reagent composition undergoes a chromogenic change that is independent of an amount of hematocrit in the whole blood, and wherein the test pad is examined for the chromogenic change visually or by instrument.

7. The analyte test device of claim 2 wherein the carrier matrix of the filter pad is selected from the group consisting of silica gel, alumina, diatomaceous earth, filter paper, chromatographic paper, glass fiber, cellulose acetate, polyvinyl chloride, polyacrylamide, polyacrylate, polyurethane, crosslinked dextran, polyethylene, polypropylene, polyvinylidene fluoride, polysulfone, and combinations thereof.

8. The analyte test device of claim 2 wherein the agglutinin is a lectin selected from the group consisting of Abrus precatorius (abrin, Jequirty bean), Agaricus bisporus (mushroom), Bauhinia purpurea (camels foot tree), Caragana arborescens (Siberian pea tree), Cicer arietinum (chick pea), Codium fragile (Green marine algae), Canavalia ensiformis (Con A, Concanavalin A, Jack bean), Datura stramonium (jimson weed), Glycine max (Soybean), Lathyrus odoratus (Sweet Pea), Lens culinaris (Lentil), Limulus polyphemus (Horseshoe crab, limulin), Lycopersicon esculentum (Tomato), Maclura pomifera (Osage orange), Mycoplasma gallisepticum, Naja mocambique mocambique (cobra venom), Naja naja kaouthia (cobra venom), Perseau americana (Avocado), Phaseolus coccineus (Scarlet runner bean), Phaseolus vulgaris (Red Kidney bean), Phytolacca americana (Pokeweed), Pisum sativum (garden pea), Pseudomonas aeruginosa, Psophocarpus tetragonolobus (winged bean), Ricinus communis (Castor bean), Robinia pseudoacacia (black locust, false acacia), Sambucus nigra (elder), Solanum tuberosum (Potato), Triticum vulgaris (Wheat germ), Vicia faba (fava bean, broad bean), Vicia sativa, Vigna radiata (Mung bean), Viscum album (European mistletoe), Wisteria floribunda (Japanese wisteria), and combinations thereof.

9. The analyte test device of claim 2 wherein the coagulant is a thrombin compound.

10. The analyte test device of claim 9 wherein the thrombin compound is selected from the group consisting of bovine thrombin, human thrombin, horse thrombin, goat thrombin, mouse thrombin, rat thrombin, pig thrombin, sheep thrombin and combinations thereof.

11. The analyte test device of claim 9 wherein the thrombin compound is selected from the group consisting of acutase, agkistrodon contortrix, ancrod, atroxin, crotalase and combinations thereof.

12. A method of manufacturing a filter pad for separating cellular components from a whole blood sample said method comprising the steps of:

a) forming a solution of a separating reagent composition, said separating reagent composition comprising;

i) from about 4 to about 4000 units of an agglutinin, or from about 90 to about 900 NIH units of a coagulant, or a combination thereof,

ii) from about 0.05% to about 3% by weight of a nonhemolytic surfactant, and

iii) an aqueous carrier;

b) applying the separating reagent composition solution to a single carrier matrix; and

c) removing a sufficient amount of the aqueous carrier from the separating reagent composition solution applied to the carrier matrix to dry the carrier matrix, and provide the filter pad comprising the single carrier matrix having homogeneously incorporated therein from about 5 to about 50 units of the agglutinin per cm³ of the carrier matrix or from about 40 to about 100 NIH units of the coagulant per cm³ of the carrier matrix, or a combination thereof; and

from about 0.3% to about 1.5% by weight of the carrier matrix of the nonhemolytic surfactant, wherein the nonhemolytic surfactant is selected from the group consisting of octoxynol 40, PEG-36 castor oil, C_11-15 pareth-12, C_12-15 pareth-9, ceteareth-27, ceteth-30, laureth-23, oleth-15, PEG-20 laurate, PEG-36 oleate, PEG-44 sorbitan laurate, PEG-30 stearate, steareth-27, PEG-11 cocamide, PEG-15 cocamine, PEG-15 tallow amine, PEG-20 hydrogenated castor oil, PPG-5-ceteth 20, PPG-myreth-11, steareth-10, trideceth-15, C_11-15 pareth-7 carboxylic acid, C_12-13 pareth-5 carboxylic acid, C_12-15 pareth-7 carboxylic acid, ceteareth-25 carboxylic acid, deceth-7 carboxylic acid, isosteareth-6 carboxylic acid, isosteareth-11 carboxylic acid, coceth-7 carboxylic acid, laureth-5 carboxylic acid, laureth-10 carboxylic acid, trideceth-4 carboxylic acid, trideceth-7 carboxylic acid, trideceth-15 carboxylic acid, trideceth-19 carboxylic acid, C_12-15 pareth-2 phosphate, ceteareth-4 phosphate, deceth-4 phosphate, dilaureth-10 phosphate, dioleth-8 phosphate, laneth-4 phosphate, laureth-3 phosphate, laureth-8 phosphate, nonoxynol-6 phosphate, nonoxynol-9 phosphate, nonoxynol-10 phosphate, nonyl nonoxynol-7 phosphate, nonyl nonoxynol-9 phosphate, nonyl nonoxynol-10 phosphate, nonyl nonoxynol-15 phosphate, nonyl nonoxynol-24 phosphate, oleth-3 phosphate, oleth-4 phosphate, oleth-10 phosphate, oleth-20 phosphate, potassium octoxynol-12 phosphate, PPG-5-ceteth phosphate, PPG-10 cetyl ether phosphate, triceteareth-4 phosphate, triceteth-5 phosphate, trideceth-6 phosphate, trilaneth-4 phosphate, trilaureth-4 phosphate, trioleth-8 phosphate, and combinations thereof,

wherein the amount of agglutinin or coagulant incorporated in the carrier matrix is about two to about five times less than the amount of agglutinin or coagulant used in the absence of a nonhemolytic surfactant.

13. An analyte test device that separates undiluted plasma or serum from an undiluted whole blood sample and that determines the presence or concentration of a soluble component of the undiluted plasma or serum, said test device consisting essentially of:

a support strip;

a test pad secured to the support strip, the test pad comprising a substrate material incorporating an indicator reagent composition that undergoes a detectable change in response to the soluble component of the undiluted plasma or serum when in contact with the undiluted plasma or serum; and

a filter pad of sufficient size to separate cellular and particulate components of the undiluted whole blood sample from the undiluted plasma or serum, the filter pad consisting essentially of a single carrier matrix having homogeneously incorporated therein from about 5 to about 50 units of an agglutinin per cm³ of the carrier matrix, and from about 0.3% to about 1.5% by weight of the carrier matrix of a nonhemolytic surfactant selected from the group consisting of octoxynol 40, PEG-36 castor oil, C_11-15 pareth-12, C_12-15 pareth-9, ceteareth-27, ceteth-30, laureth-23, oleth-15, PEG-20 laurate, PEG-36 oleate, PEG-44 sorbitan laurate, PEG-30 stearate, steareth-27, PEG-11 cocamide, PEG-15 cocamine, PEG-15 tallow amine, PEG-20 hydrogenated castor oil, PPG-5-ceteth 20, PPG-myreth-11, steareth-10, trideceth-15, C_11-15 pareth-7 carboxylic acid, C_12-13 pareth-5 carboxylic acid, C_12-15 pareth-7 carboxylic acid, ceteareth-25 carboxylic acid, deceth-7 carboxylic acid, isosteareth-6 carboxylic acid, isosteareth-11 carboxylic acid, coceth-7 carboxylic acid, laureth-5 carboxylic acid, laureth-10 carboxylic acid, trideceth-4 carboxylic acid, trideceth-7 carboxylic acid, trideceth-15 carboxylic acid, trideceth-19 carboxylic acid, C_12-15 pareth-2 phosphate, ceteareth-4 phosphate, deceth-4 phosphate, dilaureth-10 phosphate, dioleth-8 phosphate, laneth-4 phosphate, laureth-3 phosphate, laureth-8 phosphate, nonoxynol-6 phosphate, nonoxynol-9 phosphate, nonoxynol-10 phosphate, nonyl nonoxynol-7 phosphate, nonyl nonoxynol-9 phosphate, nonyl nonoxynol-10 phosphate, nonyl nonoxynol-15 phosphate, nonyl nonoxynol-24 phosphate, oleth-3 phosphate, oleth-4 phosphate, oleth-10 phosphate, oleth-20 phosphate, potassium octoxynol-12 phosphate, PPG-5-ceteth phosphate, PPG-10 cetyl ether phosphate, triceteareth-4 phosphate, triceteth-5 phosphate, trideceth-6 phosphate, trilaneth-4 phosphate, trilaureth-4 phosphate, trioleth-8 phosphate, and combinations thereof;

wherein the filter pad is positioned such that the undiluted whole blood sample first contacts the filter pad to chromatographically flow through the filter pad to contact the test pad, and

wherein the amount of agglutinin or coagulant incorporated in the carrier matrix is about two to about five times less than the amount of agglutinin or coagulant used in the absence of a nonhemolytic surfactant.

14. The analyte test device of claim 13 wherein the carrier matrix of the filter pad is filter paper.

15. The analyte test device of claim 13 wherein the nonhemolytic surfactant is selected from the group consisting of PEG-36 castor oil, PPG-5-ceteth-10 phosphate, and combinations thereof.

16. An analyte test device that separates undiluted plasma or serum from an undiluted whole blood sample and that determines the presence or concentration of a soluble component of the undiluted plasma or serum, said test device consisting essentially of:

a support strip;

a test pad secured to the support strip, the test pad comprising a substrate material incorporating an indicator reagent composition that undergoes a detectable change in response to the soluble component of the undiluted whole blood sample when in contact with the undiluted plasma or serum; and

a filter pad of sufficient size to separate the cellular and particulate components of the undiluted whole blood sample from the undiluted plasma or serum, the filter pad consisting essentially of a single carrier matrix having homogeneously incorporated therein from about 40 to about 100 NIH units of a coagulant per cm³ of the carrier matrix, or a mixture thereof, and from about 0.3% to about 1.5% by weight of the carrier matrix of a nonhemolytic surfactant is selected from the group consisting of octoxynol 40, PEG-36 castor oil, C_11-15 pareth-12, C_12-15 pareth-9, ceteareth-27, ceteth-30, laureth-23, oleth-15, PEG-20 laurate, PEG-36 oleate, PEG-44 sorbitan laurate, PEG-30 stearate, steareth-27, PEG-11 cocamide, PEG-15 cocamine, PEG-15 tallow amine, PEG-20 hydrogenated castor oil, PPG-5-ceteth 20, PPG-myreth-11, steareth-10, trideceth-15, C_11-15 pareth-7 carboxylic acid, C_12-13 pareth-5 carboxylic acid, C_12-15 pareth-7 carboxylic acid, ceteareth-25 carboxylic acid, deceth-7 carboxylic acid, isosteareth-6 carboxylic acid, isosteareth-11 carboxylic acid, coceth-7 carboxylic acid, laureth-5 carboxylic acid, laureth-10 carboxylic acid, trideceth-4 carboxylic acid, trideceth-7 carboxylic acid, trideceth-15 carboxylic acid, trideceth-19 carboxylic acid, C_12-15 pareth-2 phosphate, ceteareth-4 phosphate, deceth-4 phosphate, dilaureth-10 phosphate, dioleth-8 phosphate, laneth-4 phosphate, laureth-3 phosphate, laureth-8 phosphate, nonoxynol-6 phosphate, nonoxynol-9 phosphate, nonoxynol-10 phosphate, nonyl nonoxynol-7 phosphate, nonyl nonoxynol-9 phosphate, nonyl nonoxynol-10 phosphate, nonyl nonoxynol-15 phosphate, nonyl nonoxynol-24 phosphate, oleth-3 phosphate, oleth-4 phosphate, oleth-10 phosphate, oleth-20 phosphate, potassium octoxynol-12 phosphate, PPG-5-ceteth phosphate, PPG-10 cetyl ether phosphate, triceteareth-4 phosphate, triceteth-5 phosphate, trideceth-6 phosphate, trilaneth-4 phosphate, trilaureth-4 phosphate, trioleth-8 phosphate, and combinations thereof;

wherein the filter pad is positioned such that the undiluted whole blood sample first contacts the filter pad to chromatographically flow through the filter pad to contact the test pad, and

wherein the amount of agglutinin or coagulant incorporated in the carrier matrix is about two to about five times less than the amount of agglutinin or coagulant used in the absence of a nonhemolytic surfactant.