Characterization of Monoclonal Antibody N-Glycan Conformations by Novel Hydrophilic Interaction Liquid Chromatography (HILIC) with Trapped Ion Mobility Mass Spectrometry (TIMS) and Fluorescence Detection (FLD)
ANALYTICAL LETTERS, cilt.58, sa.12, ss.2158-2173, 2025 (SCI-Expanded, Scopus)
- Yayın Türü: Makale / Tam Makale
- Cilt numarası: 58 Sayı: 12
- Basım Tarihi: 2025
- Doi Numarası: 10.1080/00032719.2024.2384700
- Dergi Adı: ANALYTICAL LETTERS
- Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, Aerospace Database, Aquatic Science & Fisheries Abstracts (ASFA), BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, Chemical Abstracts Core, Chimica, Communication Abstracts, Food Science & Technology Abstracts, Metadex, Veterinary Science Database, Civil Engineering Abstracts
- Sayfa Sayıları: ss.2158-2173
- Anahtar Kelimeler: High-performance liquid chromatography (HPLC), hydrophilic interaction liquid chromatography with fluorescence detection (HILIC-FLD), monoclonal antibodies, N-glycan, trapped ion-mobility mass spectrometry (TIMS)
- Hacettepe Üniversitesi Adresli: Evet
Özet
Monoclonal antibodies (mAbs) have become prevalent in the pharmaceutical sector for treating various diseases and have a significant market presence. The determination of these molecules is complex and challenging. It is necessary to employ high-throughput technologies to characterize these pharmaceuticals in order to characterize sequencing, structure, composition, conformation, and mass. It is important to perform an N-glycosylation study on these macromolecules as their N-glycan profiles have a significant impact on their effectiveness. However, the conformational features of the N-glycans on mAb are not adequately addressed in commonly used methods. This study incorporated ion-mobility mass spectrometry as an additional dimension to established hydrophilic liquid chromatography trapped ion mobility spectrometry with fluorescence detection ((HILIC)LC/TIMS/FLD) in order to enhance the characterization of N-glycan structures. The N-glycans derived from two distinct mAbs and an immunoglobulin G (IgG) protein were labeled with procainamide and determined by (HILIC)LC/FLD with TIMS. The N-glycan profiles obtained from mAbs and IgG were examined in terms of conformation. The alterations in the N-glycan conformations were ascertained with the insertion of distinct monosaccharide units, such as galactose, into the structure. This method can be employed to clarify the intricate structures of N-glycans and offer insights into the conformational features of monoclonal antibodies throughout their production.