Monocyte activation test better able to detect non-endotoxin pyrogenic contaminants in medical products
First Claim
1. A method of detecting non-endotoxin pyrogens in a sample comprising the steps of:
- combining a monocyte-containing reagent and the sample to be tested in a first assay system which includes at least one microtiter well shaped such that the monocyte-containing reagent is concentrated to provide greater cell to cell contact as compared to a flat-bottomed microtiter well, wherein the surface of the microtiter well comprises a polypropylene coating or the entire microtiter well is composed of polypropylene;
incubating the monocyte-containing reagent and the sample, wherein, when the sample comprises a pyrogen, the monocyte-containing reagent produces a cytokine or an endogenous mediator of the inflammatory response;
transferring the contents of the first assay system to a second assay system which comprises at least one surface treated with an antibody to the cytokine or endogenous mediator of the inflammatory response; and
assaying the second assay system for the presence of cytokine or endogenous mediator bound to the antibody on the surface;
whereby an elevated level of cytokine or endogenous mediator bound to the surface indicates the presence of the pyrogens in the sample tested.
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Accused Products
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.
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Citations
70 Claims
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1. A method of detecting non-endotoxin pyrogens in a sample comprising the steps of:
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combining a monocyte-containing reagent and the sample to be tested in a first assay system which includes at least one microtiter well shaped such that the monocyte-containing reagent is concentrated to provide greater cell to cell contact as compared to a flat-bottomed microtiter well, wherein the surface of the microtiter well comprises a polypropylene coating or the entire microtiter well is composed of polypropylene; incubating the monocyte-containing reagent and the sample, wherein, when the sample comprises a pyrogen, the monocyte-containing reagent produces a cytokine or an endogenous mediator of the inflammatory response; transferring the contents of the first assay system to a second assay system which comprises at least one surface treated with an antibody to the cytokine or endogenous mediator of the inflammatory response; and assaying the second assay system for the presence of cytokine or endogenous mediator bound to the antibody on the surface;
whereby an elevated level of cytokine or endogenous mediator bound to the surface indicates the presence of the pyrogens in the sample tested. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35)
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36. A method of detecting non-endotoxin pyrogens in a sample comprising the steps of:
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combining a monocyte-containing reagent and the sample to be tested in an assay system comprising (i) at least one microtiter well shaped such that the monocyte-containing reagent is concentrated to provide greater cell to cell contact as compared to a flat-bottomed microtiter well, wherein the surface of the microtiter well comprises a polypropylene coating or the entire microtiter well is composed of polypropylene, and (ii) at least one surface treated with an antibody to a cytokine or an endogenous mediator of the inflammatory response; incubating the monocyte-containing reagent and the sample, wherein, when the sample comprises a pyrogen, the monocyte-containing reagent produces a cytokine or an endogenous mediator of the inflammatory response, and assaying the assay system for the presence of cytokine or endogenous mediator bound to the antibody on the surface;
whereby an elevated level of cytokine or endogenous mediator bound to the surface indicates the presence of the pyrogens in the sample tested. - View Dependent Claims (37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70)
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Specification