Claims
- 1. A method of detecting pyrogens in a sample comprising the steps of:
a) combining a pyrogen-free monocyte-containing reagent and the sample to be tested in a pyrogen-free assay system, wherein the assay system comprises at least one surface treated with a pyrogen-free antibody to a cytokine; and b) assaying the assay system for the presence of cytokine bound to the antibody on the surface; whereby an elevated level of cytokine bound to the surface indicates the presence of pyrogens in the sample tested.
- 2. The method of claim 1 wherein the monocyte-containing reagent comprises whole blood.
- 3. The method of claim 2 wherein the monocyte-containing reagent further comprises a diluent.
- 4. The method of claim 2 wherein the monocyte-containing reagent further comprises an anticoagulant.
- 5. The method of claim 4 wherein the monocyte-containing reagent further comprises a diluent.
- 6. The method of claim 3 wherein the diluent is RPMI media.
- 7. The method of claim 2 wherein the whole blood is human whole blood.
- 8. The method of claim 1 wherein the cytokine is selected from the group consisting of interleukin-1, interleukin-1ra, interleukin-6, interleukin-8, tumor necrosis factor-α, and prostaglandin E2.
- 9. The method of claim 8 wherein the cytokine is interleukin-6.
- 10. The method of claim 1 wherein the assay system includes at least one microtiter well, and the surface on which the antibody is applied is a portion of the interior of the microtiter well.
- 11. The method of claim 10 wherein the microtiter well is made from polystyrene or polypropylene.
- 12. The method of claim 1 wherein the assay system is assayed by a colorimetric enzyme-linked immunosorbent assay for the cytokine.
- 13. The method of claim 1 wherein the assay system is assayed by a radio-labeled immunoassay for the cytokine.
- 14. The method of claim 1 wherein the assay system is assayed by a fluorescence-labeled immunoassay for the cytokine.
- 15. The method of claim 1 wherein the pyrogen-free antibody is a polyclonal antibody.
- 16. The method of claim 1 wherein the pyrogen-free antibody is a monoclonal antibody.
- 17. The method of claim 1 wherein the sample is a medicinal drug for injection or infusion.
- 18. The method of claim 1 wherein the sample is a blood product.
- 19. A pyrogen-free assay system wherein the assay system comprises a surface coated with a pyrogen-free antibody to a cytokine.
- 20. The pyrogen-free assay system of claim 19 wherein the assay system further comprises at least one microtiter well, and wherein the surface to which the pyrogen-free antibody is applied is a portion of the interior of the microtiter well.
- 21. The pyrogen-free assay system of claim 20 wherein the well is made from polystyrene or polypropylene.
- 22. The pyrogen-free assay system of claim 20 wherein the well is part of a planar array of similar wells, so situated so that the array of wells may be read with automated reading equipment.
- 23. The pyrogen-free assay system of claim 19 wherein the cytokine is selected from the group consisting of interleukin-1, interleukin-1ra, interleukin-6, interleukin-8, tumor necrosis factors-α, and prostaglandin E2.
- 24. The pyrogen-free assay system of claim 19 wherein the cytokine is interleukin-6.
- 25. The pyrogen-free assay system of claim 19 wherein the pyrogen-free antibody is a polyclonal antibody.
- 26. The pyrogen-free assay system of claim 19 wherein the pyrogen-free antibody is a monoclonal antibody.
- 27. A method of detecting pyrogens in a sample comprising the steps of:
a) combining a pyrogen-free monocyte-containing reagent and the sample to be tested in a pyrogen-free assay system, wherein the assay system comprises at least one surface treated with a pyrogen-free antibody to an endogenous mediator of the inflammatory response; and b) assaying the assay system for the presence of the mediator bound to the antibody on the surface; whereby an elevated level of the mediator bound to the surface indicates the presence of pyrogens in the sample tested.
- 28. The method of claim 27 wherein the endogenous mediator of the inflammatory response is endothelin.
- 29. A method of measuring the capacity of a leukocyte to respond to a stimuli comprising the steps of:
a) combining a pyrogen-free leukocyte-containing reagent and a stimuli in a pyrogen-free assay system, wherein the assay system comprises at least one surface treated with a pyrogen-free antibody to an endogenous mediator of the inflammatory response; and b) assaying the assay system for the presence of the mediator bound to the antibody on the surface; whereby an elevated level of the mediator bound to the surface indicates the response of the leukocyte to the stimuli.
- 30. The method of claim 29 wherein the stimuli is selected from the group consisting of endotoxin, an infectious agent, vaccine, adjuvant, and drug.
- 31. The method of claim 29 wherein the leukocyte-containing reagent comprises whole human blood.
- 32. The method of claim 29 wherein the endogenous mediator of the inflammatory response comprises a cytokine.
- 33. A method of measuring the basal production of endogenous mediators of the inflammatory response by leukocytes comprising the steps of:
a) adding a pyrogen-free leukocyte-containing reagent to a pyrogen-free assay system, wherein the assay system comprises at least one surface treated with a pyrogen-free antibody to an endogenous mediator of the inflammatory response; and b) assaying the assay system for the presence of the mediator bound to the antibody on the surface; whereby the amount of mediator bound to the surface indicates the basal production of endogenous mediators of the inflammatory response by the leukocytes.
- 34. The method of claim 33 wherein the leukocyte-containing reagent comprises whole human blood.
- 35. The method of claim 33 wherein the endogenous mediator of the inflammatory response comprises a cytokine.
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application claims priority from Provisional Application Ser. No. 60/168,972 filed on Dec. 3, 1999, which is hereby incorporated by reference in its entirety.
Provisional Applications (1)
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Number |
Date |
Country |
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60168972 |
Dec 1999 |
US |