Monocyte activation test better able to detect non-endotoxin pyrogenic contaminants in medical products

• US 9,023,647 B2
• Filed: 11/10/2011
• Issued: 05/05/2015
• Est. Priority Date: 12/22/2005
• Status: Active Grant

• Current Assignee: Baxter Healthcare SA (Baxter International Inc.), Baxter International Inc., Secretary of State for Health
• Original Assignee: Baxter Healthcare SA (Baxter International Inc.), Baxter International Inc., Secretary of State for Health
• Inventors: Poole, Stephen, Patel, Mehul
• Primary Examiner(s): STANFIELD, CHERIE MICHELLE
• Application Number:
  US13/293,868
• Publication Number:
  US 20120058559A1
• Time in Patent Office: 1,272 Days
• Field of Search: 435/383, 435/305.2
• US Class Current: 435/383
• CPC Class Codes: G01N 2333/5412 IL-6
• CPC Class Codes: G01N 2800/7095 Inflammation
• CPC Class Codes: G01N 33/5047 Cells of the immune system
• CPC Class Codes: G01N 33/5055 involving macrophages
• CPC Class Codes: G01N 33/5091 for testing the pathological state of an organism
• CPC Class Codes: G01N 33/6863 Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
• CPC Class Codes: G01N 33/6869 Interleukin

Abstract

An improved monocyte activation test is described that is better able to detect non-endotoxin pyrogens in medical products, in which a sample is incubated with a monocyte-containing reagent in an assay system comprising at least one surface comprising polypropylene. The invention also concerns assay systems for use in these tests that include at least one microtiter well having at least one interior surface comprising polypropylene and having a shape such that monocyte-containing reagent is concentrated in the well to provide greater cell to cell contact. The invention also relates to a diagnostic kit that can be used to test for the presence of non-endotoxin pyrogens in a sample.

Claims

1. A diagnostic kit comprising:

(i) a microtiter plate comprising a plurality of microtiter wells shaped such that, when cells are contained in each well, the cells are concentrated, as compared to a flat-bottomed microtiter well, wherein the surface of each well comprises a polypropylene coating or is composed of polypropylene; and

(ii) a cryopreserved monocyte-containing reagent, wherein the cryopreserved monocyte-containing reagent is provided to an end-user separately from the microtiter plate.

2. The diagnostic kit of claim 1, wherein the cryopreserved monocyte-containing reagent is provided to the end-user by shipping on dry ice.

3. The diagnostic kit of claim 1, wherein the cyropreserved monocyte-containing reagent is provided to an end-user aliquoted into the optimum number of cells per well.

4. The diagnostic kit of claim 1, wherein the cryopreserved monocyte-containing reagent comprises cryopreserved peripheral blood mononuclear cells, cryopreserved monocytic cell line cells, or cryopreserved whole blood.

5. The diagnostic kit of claim 4, wherein the monocytic cell line comprises a human monocytic cell line selected from the group consisting of:

MONOMAC-6, THP-1, 28SC, and a cell line having endogenous CD14 and Toll-like receptors or which have been transfected with CD14 and/or Toll-like receptors and/or reporter genes for inflammatory or pyrogenic mediators.

6. The diagnostic kit of claim 1, further comprising an antibody to a cytokine or an endogenous mediator of the inflammatory response.

7. The diagnostic kit of claim 6, wherein the cytokine or endogenous mediator of the inflammatory response is selected from the group consisting of interleukin-1 (IL-1), interleukin-1ra (IL-1ra), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-α

, (TNF-α

), and endothelin.

8. The diagnostic kit of claim 6, wherein at least one surface of the microtiter wells of the microtiter plate is treated with the antibody.

9. The diagnostic kit of claim 6, further comprising a second microtiter plate comprising at least one surface treated with the antibody.

10. The diagnostic kit of claim 1, wherein each microtiter well in the plurality comprises an open top, an upper region extending downwardly from the open top, and a bottom wall tapering in diameter from a location above the lowest point to the lowest point.

11. The diagnostic kit of claim 1, wherein each microtiter well in the plurality comprises an open top, an upper region extending downwardly from the open top having a top end and a bottom end, and a bottom region having a top end and a lowest point, the bottom region extending from the bottom end of the upper region and tapering in diameter more rapidly than the upper region from the top end of the bottom region toward the lowest point.

12. The diagnostic kit of claim 1, wherein the bottom wall of each microtiter well in the plurality is non-planar, curved, parabolic or downwardly extending, or the sides of the microtiter well are sloped inward.