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Updated: Aug 10, 2026
IP-FCM: Immunoprecipitation Detected by Flow Cytometry
Published on: December 2, 2010
Flow-cytometric method for the quantitation of the Fc function of intravenous immunoglobulin preparations
I Ramasamy1, E Tran, A Charnock
1Molecular Biology Section, Therapeutic Goods Administration Laboratories, Woden, Australia. indra.ramasamy@health.gov.au
Objectives:
We have developed and optimised a new flow-cytometric method for the measurement of the Fc function of intravenous immunoglobulin (IVIg) 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 Fc gammaRI and Fc gammaRII 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.
