Measurement of isoagglutinins in immunoglobulins for intravenous application by flow cytometry.
Journal
Analytical biochemistry
ISSN: 1096-0309
Titre abrégé: Anal Biochem
Pays: United States
ID NLM: 0370535
Informations de publication
Date de publication:
15 02 2020
15 02 2020
Historique:
received:
19
08
2019
revised:
21
11
2019
accepted:
05
12
2019
pubmed:
11
12
2019
medline:
20
11
2020
entrez:
11
12
2019
Statut:
ppublish
Résumé
Isoagglutinins present in intravenous immunoglobulin (IVIG) products have been linked to haemolysis. Therefore, accurately assessing isoagglutinin content in IVIG products is important. The standard European Pharmacopoeia (Ph.Eur.) direct assay is limited by low precision. Here, we describe the development of a fluorescence-activated cell sorting (FACS) method for assessing isoagglutinin levels. Serially diluted IVIG samples were incubated with red blood cells (RBCs), RBC-bound anti-A and anti-B antibodies were detected using a fluorescently-labelled antibody and the median fluorescence intensity of samples was assessed by FACS. Results were compared with the Ph.Eur. direct assay. The method was used to determine isoagglutinins in commercial products produced with and without isoagglutinin reduction steps. Assay precision, reported as the coefficient of variation, for the FACS method was 14% and 8% for anti-A and anti-B, respectively versus 33% and 20% with the Ph.Eur. direct assay. Application of the method on commercially available IVIGs revealed differences in isoagglutinin content between products produced with and without isoagglutinin reduction steps. This FACS assay allows for quantification of isoagglutinin concentrations in IVIGs with higher precision than the Ph.Eur. direct assay. Also the FACS assay confirms differences in isoagglutinin levels between IVIG products and the efficacy of isoagglutinin reduction measures.
Identifiants
pubmed: 31821802
pii: S0003-2697(19)30774-2
doi: 10.1016/j.ab.2019.113534
pii:
doi:
Substances chimiques
Hemagglutinins
0
Immunoglobulins, Intravenous
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
113534Informations de copyright
Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of competing interest All authors are currently, or were previously, employed by CSL Behring, the funders of this research. In addition, Alphonse Hubsch and Dominik Stadler own shares of CSL. There are no further conflicts of interest to report.