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US 5,176,998 AGranted
Cell surface antigen detection method
AssigneeThe Scripps Research Institute
InventorRobert McMillan
Life of this patent
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Abstract
The present invention provides a method for assaying a sample of cells for the presence of a cell surface antigen. More particularly, the present invention describes methods for detecting the presence of plasma and cell bound autoantibodies against cell surface antigens. In addition, the present invention provides a method of crossmatching donor platelets and transfusion recipients.
Claims9
A method of assaying a sample of cells for the presence of a cell surface antigen comprising:
- (a) admixing said cells with an antibody capable of immunoreacting with said antigen to form a cell-sensitizing reaction admixture,
- (b) maintaining said cell-sensitizing reaction admixture for a time period sufficient to form an immunocomplex containing said antigen and said antibody, thereby forming sensitized cells,
- (c) lysing said sensitized cells to provide an aqueous composition, said composition containing a solubilized immunocomplex when said immunocomplex is formed in step (b),
- (d) admixing said aqueous composition with a solid support capable of specifically binding said solubilized immunocomplex, thereby forming a solid/liquid phase admixture,
- (e) maintaining said solid/liquid phase admixture for a time period sufficient for any of said solubilized immunocomplex present to bind to said solid support, thereby forming a solid phase-bound complex, and
- (f) assaying for the presence of any bound complex that formed in step (e) and thereby for the presence of said cell surface antigen, by reacting the bound complex present with a labeled specific binding agent that specifically binds to the immunocomplex portion of the bound complex.
3 more elements
A method of assaying for a platelet surface antigen-antibody complex in a sample of platelets comprising the steps of:
- (a) lysing said sample of paltelets to provide an aqueous composition containing a solubilized platelet surface antigen-antibody complex,
- (b) forming a capturing reaction admixture by admixing said aqueous composition with a solid support capable of immunoreacting with said solubilized platelet surface antige-antibody complex,
- (c) maintaining said capturing reaction admixture for a time period sufficient for said solid support to immunoreact with any of said solubilized platelet surface antigen-antibody complex present to form a solid phase-affixed immunoreaction product, said maintaining thereby forming a maintained solid support,
- (d) separating the maintained solid support from any non-bound protein,
- (e) forming a labeling reaction admixture by admixing said maintained solid support with a labeled antibody molecule capable of immunoreacting with said antigen-antibody complex of said solid phase-affixed immunoreaction product,
- (f) maintaining said labeling reaction admixture for a time period sufficient for said labeled antibody molecule to immunoreact with any of said solid phase-affixed immunoreaction product formed in step (c) to form a labeled solid-phase complex, and
- (g) assaying for the presence of any labeled solid phase-affixed complex formed in step (e), and thereby for the presence of any platelet surface antigen-antibody complex in said sample of platelets.
4 more elements
A method of assaying sample of platelets for a platelet surface antigen comprising the steps of:
- (a) forming a platelet sensitizing reaction admixture by admixing a sample of platelets with an antibody molecule capable of immunoreacting with said platelet surface antigen to produce a platelet immunocomplex.
- (b) maintaining said sensitizing reaction admixture for a time period sufficient for formation of said platelet immunocomplex, thereby forming antibody-sensitized platelets.
- (c) lysing said sensitized platelets to provide an aqueous composition, said composition containing a solubilized platelet immunocomplex when said immunocomplex is formed in step (b),
- (d) forming a capturing reaction admixture by admixing said aqueous composition with a solid support capable of immunoreacting with said solubilized platelet immunocomplex,
- (e) maintaining said capturing reaction admixture for a time period sufficient for said solid support to immunoreact with any of said solubilized platelet immunocomplex present to form a solid phase-affixed immunoreaction product, said maintaining thereby forming a maintained solid support,
- (f) forming a labeling reaction admixture by admixing said maintained solid support with a labeled antibody molecule capable of immunoreacting with said platelet immunocomplex of said solid phase-affixed immunoreaction product,
- (g) maintaining said labeling reaction admixture for a time period sufficient for said labeled antibody molecule to immunoreact with any of said solid phase-affixed immunoreaction product formed in step (e) to form a labeled solid phase complex, and
- (h) assaying for the presence of any labeled solid phase-affixed complex formed in step (g), and thereby for the presence of said platelet surface antigen in said sample of platelets.
5 more elements
Dependencies are inferred from the claim text.