Methods of detecting antibodies specific for denatured HLA antigens

A method of screening for antibodies that specifically bind an HLA antigen in its native conformation and said HLA antigen in a denatured conformation, comprising the steps of:

• preparing a panel comprising a first solid-phase substrate having an HLA antigen in its native conformation immobilized or attached thereto and a second solid-phase substrate having said HLA antigen solely in a denatured conformation immobilized or attached thereto,
• obtaining a serum sample from a human subject,
• contacting the panel with the serum sample, and
• detecting distinguishable binding of antibodies in said serum sample to the HLA antigen in its native conformation and binding of antibodies in said serum sample to said HLA antigen in its denatured conformation, wherein the binding of the antibodies to said HLA antigens in its denatured conformation identifies the potential presence of antibodies which are specific for said HLA antigen in a denatured conformation.