Making encoded ghost cells for multiplexed detection of anti-red cell alloantibodies
AssigneeBioArray Solutions, Ltd.
InventorSukanta Banerjee
Life of this patent
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Abstract
Disclosed are methods for detecting antibody in a sample, where the antibody targets an antigen expressed by red blood cells or red blood cell ghosts. Rather than detecting the binding events between a particular antigen antibody pair (as in traditional agglutination based assays) the methods herein allow for multiplexed detection of clinically important allo-immune antibodies to blood group antigens. Specifically the method involves generating fluorescently encoded red blood cells or red blood cell ghosts with known antigen presentation and using them to detect the presence of antibody in serum/plasma with a fluorescent sandwich type immunoassay. The assay results can be read using flow cytometric or fluorescent microscope based imaging techniques.
Claims8
A method of preparing encoded ghost cells comprising:
- separating erythrocytes from whole blood;
- incubating the erythrocytes with a dye solution, wherein the dye solution contains two or more fluorescent dyes mixed at a predetermined ratio which upon excitation exhibit distinct fluorescent emission signals, wherein the ratio of the fluorescent emission signals is governed by said predetermined ratio;
- lysing the erythrocytes in the dye solution by adding a lysing solution;
- re-sealing the lysed erythrocytes by adding a re-sealing solution;
- and
- separating the re-sealed erythrocytes to give encoded ghost cells.
Dependencies are inferred from the claim text.