Journalartikel

Jahr der Veröffentlichung: 2013

Seiten: 193-201

Zeitschrift: Transfusion

Bandnummer: 53

Heftnummer: 1

ISSN: 0041-1132

DOI Link: https://doi.org/10.1111/j.1537-2995.2012.03675.x

Verlag: Wiley

Abstract: 
BACKGROUND: Several methods exist for the detection of neutrophil antibodies; most of them, however, require fresh neutrophils. In this study, an enzyme-linked immunosorbent assay (ELISA) using recombinant HNA-1 antigens (rHNAs) was developed to detect HNA-1a, -1b, and -1c alloantibodies in serum samples. STUDY DESIGN AND METHODS: Soluble rHNA-1a, -1b, and -1c were isolated from culture supernatant of transfected insect cells. Purified rHNA antigens were immobilized on microtiter wells using antibody against V5-Tag protein. Sera were added, and bound antibodies were detected by enzyme-labeled secondary antibodies. In parallel, monoclonal antibodyimmobilized granulocyte antigen (MAIGA) was performed with two different monoclonal antibodies (MoAbs) against Fc?RIIIb (3G8 and BW209). RESULTS: Fifteen MAIGA-positive sera containing HNA-1a alloantibodies were tested in ELISA. Thirteen of 15 (86.7%) MAIGA-positive sera captured by MoAbs 3G8 and/or BW209 reacted specifically with rHNA-1a. Four (26.7%) HNA-1a sera showed additional reaction with rHNA-1c. When anti-HNA-1b alloantibodies were analyzed in ELISA, 13 of 15 (86.7%) showed specific positive reaction with rHNA-1b, and 12 of 15 (80.0%) cross-reacted with rHNA-1c. Two HNA-1c sera reacted specifically with rHNA-1c. Immunoprecipitation analysis of all ELISA-negative HNA-1a and -1b sera did not show any specific band indicating false-positive reaction of these sera in MAIGA assay. CONCLUSIONS: These results suggested that rapid ELISA using recombinant neutrophil antigens may provide a valuable method for rapid screening of human alloantibodies against HNA-1a, -1b, and -1c in patients with neutropenia and in blood donors.

Harvard-Zitierstil: Werth, S., Bayat, B., Tjahjono, Y., Eskandar, J., Berghoefer, H., Bein, G., et al. (2013) Rapid enzyme-linked immunosorbent assay for the detection of antibodies against human neutrophil antigens -1a,-1b, and -1c, Transfusion, 53(1), pp. 193-201. https://doi.org/10.1111/j.1537-2995.2012.03675.x

APA-Zitierstil: Werth, S., Bayat, B., Tjahjono, Y., Eskandar, J., Berghoefer, H., Bein, G., Sachs, U., & Santoso, S. (2013). Rapid enzyme-linked immunosorbent assay for the detection of antibodies against human neutrophil antigens -1a,-1b, and -1c. Transfusion. 53(1), 193-201. https://doi.org/10.1111/j.1537-2995.2012.03675.x