Journalartikel

Jahr der Veröffentlichung: 2018

Seiten: 200-207

Zeitschrift: Transfusion

Bandnummer: 58

Heftnummer: 1

ISSN: 0041-1132

eISSN: 1537-2995

DOI Link: https://doi.org/10.1111/trf.14428

Verlag: Wiley

Abstract: 

BACKGROUNDCurrently, the gold standard for the identification of antibodies against human neutrophil antigens (HNAs) is the monoclonal antibody-immobilized granulocyte antigen (MAIGA) assay. However, the handling of this assay is laborious and therefore cumbersome for the rapid screening of neutrophil antibodies.

STUDY DESIGN AND METHODSIn this study, we simplified the performance of the conventional MAIGA procedure and approved it for the identification of anti-HNA-1 with HNA-1-typed neutrophils and stable transfected (HEK293) cell line expressing HNA-1a, -1b, and -1c.

RESULTSIn contrast to the conventional MAIGA, all working steps including the incubation of antibodies with cells, washings, cell lysis, and subsequent antibody detection could be performed on microtiter plates. This modification accelerates the work schedule of MAIGA and reduces pipetting errors. Comparison between both assay performances using neutrophils as target showed concordant results. Subsequently, stable mammalian cell lines were tested. In comparison to neutrophils, cell lines were stable for a longer period of time (>4 weeks) and results obtained with these cell lines showed better interassay precision. Analysis of different FcRIIIb capture monoclonal antibodies (MoAbs) for the MAIGA assay showed that MoAb LNK16 is superior for the detection of anti-HNA-1a, -1b, and -1c, whereas MoAb 3G8 showed false-negative results, caused by competitive inhibition of anti-HNA-1c alloantibodies.

CONCLUSIONThe modification of MAIGA and the use of transfected HEK293 cells improve the detection of anti-HNA-1 alloantibodies that may allow screening analysis in large cohort of samples.

Harvard-Zitierstil: Simtong, P., Romphruk, A., Hofmann, C., Reil, A., Sachs, U. and Santoso, S. (2018) Improvement of monoclonal antibody-immobilized granulocyte antigen assay for the detection of anti-HNA-1 alloantibodies, Transfusion, 58(1), pp. 200-207. https://doi.org/10.1111/trf.14428

APA-Zitierstil: Simtong, P., Romphruk, A., Hofmann, C., Reil, A., Sachs, U., & Santoso, S. (2018). Improvement of monoclonal antibody-immobilized granulocyte antigen assay for the detection of anti-HNA-1 alloantibodies. Transfusion. 58(1), 200-207. https://doi.org/10.1111/trf.14428