Methods of detecting alloantibodies using HLA and non-HLA antigens

A method for determining the presence of reactive antibodies in serum of a human subject, said method comprising:

• a. incubating the serum with:
• a first collection of solid-phase substrates comprising at least two substrates, wherein each of the at least two substrates in the first collection comprises a different Class I or Class II human leukocyte antigen (HLA) antigen, and
• a second collection of solid-phase substrates comprising at least five substrates, wherein each of the at least five substrates in the second collection comprises a different non-HLA antigen selected from the group consisting of:
• regenerating islet-derived protein 3-alpha (Reg3a), platelet glycoprotein 4 (CD36), proteosome subunit alpha type-4 (PSMA4), angiotensinogen, tissue factor (F3), 60 kilodalton SS-A/Ro ribonucleoprotein (TROVE2), interferon-induced helicase C domain-containing protein A (IFIHI), chromatin assembly factor 1 subunit B (CHAF 1B), glutamate decarboxylase 2 (GAD2), heterogenous nuclear ribonucleoprotein K (HNRNPK), gamma-interferon inducible protein 16 (IFI16), proliferation-associated protein 2G4 (PA2G4), bactericidal/permeability-increasing protein (BPI) fold-containing family A member 1 (PLUNC), 26S protease regulatory subunit 6B (PSMC4), fibronectin leucine-rich repeat transmembrane protein 2 (FLRT2), PRKR-interacting protein 1 (PRKRIP1), endothelin receptor type A (EDNRA), alpha-enolase (ENO-1), and lamin-A,
• wherein the first collection of solid-phase substrates and the second collection of solid-phase substrates are incubated with the serum for a sufficient time for one or more antibodies specific to the Class I and/or Class II HLA antigen(s) in said serum to bind to said HLA antigen(s) on the first collection of solid-phase substrates to form one or more first complex(es), and one or more antibodies specific for the non-HLA antigen(s) in said serum to bind to said non-HLA antigen(s) on the second collection of solid-phase substrates to form one or more second complex(es), and
• b. detecting the presence of the one or more first complex(es) and the one or more second complex(es) to determine the presence of the reactive antibodies by detecting a label bound to the one or more first complex(es) and a label bound to the one or more second complex(es).