Skip to main content
Skip to main menu Skip to spotlight region Skip to secondary region Skip to UGA region Skip to Tertiary region Skip to Quaternary region Skip to unit footer


Characterization of Glycation and Other PTMs in Biotherapeutics Using HILIC-MS

Photo of Sonal Priya, speaker
Sonal Priya
Graduate Student, Department of Chemistry
University of Georgia
iSTEM Building 2, Room 1218
Analytical Seminar

Glycation is an important post-translational modification that is linked to diabetes, cataract, Alzheimer’s, Rheumatoid arthritis and Parkinson’s disease.1 This reaction occurs between a reducing sugar and a primary amine at the N-terminus of protein or the sidechain of Lysine residues.  Ultimately this interaction leads to advanced glycation end products (AGEs) that are associated with several disease complications.2 Glycation could also occur during the manufacturing and storage of therapeutic proteins, including mAbs, necessitating the characterization and accurate quantitation of this modification to ensure safety and efficacy of therapeutic drug products.3 This work focuses on the analysis of in vitro glycated peptides and proteins. Hydrophilic Interaction Chromatography (HILIC) that has been used to characterize hydrophilic modifications4 is utilized in this analysis as the hydrophilic nature of the glycation product could lead to a characteristic shift in HILIC retention. Quantitation for glycation modification has been attempted in the literature.5 Relative quantitation is currently performed by comparing the area of the glycated peptide with that of its unmodified counterparts. Accuracy of this method depends on the unmodified and modified peptides having similar ionization efficiencies. We propose the use of non-glycated SILAC-labeled proteins to provide both the relative and absolute level of glycation in human IgG1 (Adalimumab) by measuring the peak area of the unmodified peptides relative to that of the SILAC labeled protein. We expect this quantitation approach will be applicable to other PTMs.


[1] Videira, P. Q.; Castro-Caldas, M., Linking Glycation and Glycosylation With Inflammation and Mitochondrial Dysfunction in Parkinson’s Disease. Front. Neurosci. 2018, 12, 381.

[2] Kuzan, A.,Toxicity of advanced glycation end products (Review). Biomedical Reports 2021, 46.

[3] Wei, B. et. al., Glycation of antibodies: Modification, methods and potential effects on biological functions. mAbs 2017, 9, 4, 586-594.

[4] Badgett, M. J.; Boyes, B.; Orlando, R., Peptide retention prediction using hydrophilic interaction liquid chromatography coupled to mass spectrometry. J. Chromatogr. A 2018, 1537, 58-65.

[5] Li, W. et. al., State-of-the-Art and Emerging Technologies for Therapeutic Monoclonal Antibody Characterization Volume 2. Biopharmaceutical Characterization: The NISTmAb Case Study., 2015, 119-183.

Support Us

We appreciate your financial support. Your gift is important to us and helps support critical opportunities for students and faculty alike, including lectures, travel support, and any number of educational events that augment the classroom experience. Click here to learn more about giving.

Every dollar given has a direct impact upon our students and faculty.

Got More Questions?

Undergraduate inquiries: 

Registration and credit

AP Credit, Section Changes, Overrides,

Graduate inquiries:

Contact Us!

Assistant to the Department Head: Donna Spotts, 706-542-1919 

Main office phone: 706-542-1919 

Main Email:

Head of Chemistry: Prof. Jason Locklin