Send to

Choose Destination
J Immunol Methods. 2015 Feb;417:34-44. doi: 10.1016/j.jim.2014.12.004. Epub 2014 Dec 15.

A method for high-throughput, sensitive analysis of IgG Fc and Fab glycosylation by capillary electrophoresis.

Author information

Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States.
Department of Biochemistry, Oxford University, Oxford, United Kingdom.
Thayer School of Engineering, Dartmouth College, Hanover, NH, United States.
Department of Neurology, Brigham and Women's Hospital, Boston, MA, United States.
Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States; Center for Virology and Vaccine Research, Beth Israel Deaconess Medical Center, Boston, MA, United States.
Ragon Institute of MGH, MIT and Harvard, Cambridge, MA, United States. Electronic address:


The N-glycan of the IgG constant region (Fc) plays a central role in tuning and directing multiple antibody functions in vivo, including antibody-dependent cellular cytotoxicity, complement deposition, and the regulation of inflammation, among others. However, traditional methods of N-glycan analysis, including HPLC and mass spectrometry, are technically challenging and ill suited to handle the large numbers of low concentration samples analyzed in clinical or animal studies of the N-glycosylation of polyclonal IgG. Here we describe a capillary electrophoresis-based technique to analyze plasma-derived polyclonal IgG-glycosylation quickly and accurately in a cost-effective, sensitive manner that is well suited for high-throughput analyses. Additionally, because a significant fraction of polyclonal IgG is glycosylated on both Fc and Fab domains, we developed an approach to separate and analyze domain-specific glycosylation in polyclonal human, rhesus and mouse IgGs. Overall, this protocol allows for the rapid, accurate, and sensitive analysis of Fc-specific IgG glycosylation, which is critical for population-level studies of how antibody glycosylation may vary in response to vaccination or infection, and across disease states ranging from autoimmunity to cancer in both clinical and animal studies.


Capillary electrophoresis; Fc separation; Glycan analysis; IgG N-glycosylation

[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Elsevier Science Icon for PubMed Central
Loading ...
Support Center