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Flow‐Cytometric Method for the Quantitation of the Fc Function of Intravenous lmmunoglobulin Preparations
Author(s) -
Ramasamy Indra,
Tran Em,
Charnock Alison,
Farrugia Albert
Publication year - 2000
Publication title -
vox sanguinis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.68
H-Index - 83
eISSN - 1423-0410
pISSN - 0042-9007
DOI - 10.1046/j.1423-0410.2000.7830185.x
Subject(s) - dithiothreitol , flow cytometry , antibody , fragment crystallizable region , receptor , chemistry , function (biology) , fc receptor , cell culture , microbiology and biotechnology , chromatography , immunology , biochemistry , biology , enzyme , genetics
Objectives:We have developed and optimised a new flow‐cytometric method for the measurement of the Fc function of intravenous immunoglobulin (IVlg) preparations, which is important in predicting the effector function of immunoglobulin (Ig) in such preparations. Materials and Methods: Ig was bound to a monocytic cell line, THP‐1 with FcγRl and FcγRll cell surface receptors, and the bound Ig detected by FITC‐conjugated F(ab) 2 fragment of rabbit anti‐human IgG. Results: Validation studies showed that Ig bound to the cell line through the Fc portion. The method detected alterations in Fc function caused by reduction with dithiothreitol or by storage. The method was reproducible (CV<11%) and a limited comparison study showed that it correlated with the European Pharmacopoeia reference method. Conclusions: This technically simple method is suitable for the quantitation of the Fc function of Ig preparations.