Generation of human antibodies targeting human platelet antigen (HPA)-1a

This study aimed to isolate HPA-1a-specific B-cells from an HPA-1a-alloimmunized pregnant woman. Using fluorescently labeled HPA-1a-positive platelets, single B-cells were sorted and cultured for 10 days to stimulate antibody production. Subsequently, supernatants were tested for the presence of antibodies by enzyme-linked immunosorbent assay and their reactivity towards HPA-1a-positive platelets. Amplification and sequencing of variable regions allowed the generation of monoclonal antibodies using a HEK-Freestyle-based expression system.RESULTS: Three platelet-specific B-cells were obtained and cloned of which two were specific for HPA-1a, named D- and M-204, while the third was specific for HLA class I, which was named L-204.DISCUSSION: This study outlined an effective method for the isolation of HPA-1a-specific B-cells and the generation of monoclonal antibodies. Further characterization of these antibodies holds promise for better understanding the pathogenic nature of alloantibodies in FNAIT.PMID:38400657 | DOI:10.1111/trf.17758

Source: Transfusion - February 24, 2024 Category: Hematology Authors: Janita J Oosterhoff Federica Linty Remco Visser Thijs de Vos Suzanne Hofstede-van Egmond Miranda van de Weerd Leendert Porcelijn Masja de Haas Ellen van der Schoot Gestur Vidarsson Source Type: research