Method for immunobead flow cytometric detection of anti-HLA panel reactive antibody

A method for determining the percentage of panel reactive antibodies in serum of a subject against human leukocyte antigens (HLA) antigens, said method comprising:

• providing a collection of microbeads of different subtypes, wherein each subtype is coated with different purified HLA antigens, and the microbeads of at least one subtype each present HLA antigens derived from a cell population presenting the same HLA antigens;
• adding said serum from said subject to said collection of microbeads;
• incubating said serum and microbeads for sufficient time for anti-HLA antibodies in said serum to bind to said HLA antigens;
• incubating said microbeads with at least one labeled ligand capable of specifically binding with said anti-HLA antibodies bound to said HLA antigens;
• detecting the presence of labeled ligand bound to said anti-HLA antibodies by flow cytometry to determine the presence or absence of panel reactive antibodies;
• and, determining the percentage of panel reactive antibodies.