Pronase-free B-cell flow-cytometry crossmatch

Detection of anti-class II antibodies by panel response assay (PRA) and flow cross-match techniques carries an important value in terms of graft function. Even low levels of pre-formed alloantibodies to HLA class II antigens represent a risk of rejection. We present here a method for blocking non-specific flow crossmatch reactions using pooled, heat-inactivated rabbit serum. This method shows very low background and minimal non-specific reactions. In addition, it avoids the use pronase enzyme that can non-specifically digest different cell surface proteins.