An integrated stage-tip-based glycomic and glycoproteomic approach for simple and rapid N-glycosylation profiling of glycoproteins


Kayili H. M. , Ragoubi Z. M. E. , SALİH B.

BIOMEDICAL CHROMATOGRAPHY, vol.36, no.12, 2022 (SCI-Expanded) identifier identifier

  • Publication Type: Article / Article
  • Volume: 36 Issue: 12
  • Publication Date: 2022
  • Doi Number: 10.1002/bmc.5503
  • Journal Name: BIOMEDICAL CHROMATOGRAPHY
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus, BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, Chemical Abstracts Core, Chimica, EMBASE, MEDLINE, Veterinary Science Database
  • Keywords: glycomics, glycosylation, immunoglobulin G, N-glycosylation profiling, SAMPLE PREPARATION, MECHANISMS, CHROMATOGRAPHY, PROCAINAMIDE, GLYCOPEPTIDE, GLYCANS, DISEASE
  • Hacettepe University Affiliated: Yes

Abstract

Glycosylation is a post-translational modification that plays an active role in many cellular events. It also regulates many functions of proteins. Monoclonal antibody-derived drugs are used to treat many diseases, and glycosylation affects the activity of such drugs developed. On the other hand, N-glycans may change in certain diseases. Therefore, rapid and efficient bioanalytical methods are needed for N-glycosylation profiling. The study aimed to develop an integrated stage-tip application for the simple and rapid N-glycosylation profiling of glycoproteins. A fast and inexpensive N-glycosylation profiling was achieved by integrating all glycoproteomic and glycomic sample preparation steps into a stage-tip. The glycomic approach of the integrated stage-tip reduces the N-glycan profiling time from 2 days to approximately 2.5 hours. It also allows the profiling of immunoglobulin G (IgG) N-glycopeptides directly from human plasma. In addition, N-glycosylation profiling can be done in the developed method without sorbents, C-18 or others, such as strong-cation exchange, at the glycopeptide level.