In recent years, absolute quantification of target proteins has become one of the new hot topics in quantitative proteomics, but less research has been reported on the absolute quantification of glycoproteins. Recently, CD BioGlyco’s researchers have developed several strategies for absolute quantification of glycoproteins and we expect to be your scientific assistant in the field of glycoproteomics.

Introduction

Qualitative and quantitative analysis of carbohydrates conjugates is a prerequisite for the study of functional glycomics and glycobiology. Typically, high-performance anion liquid chromatography coupled with a pulsed amperometric detector, HPLC coupled with a fluorescence detector and LC-MS, are the most widely used methods for the carbohydrates analysis, providing quantitative information on the isolated glycans. Currently, quantitative techniques are increasingly employed in the field of glycomics and glycoproteomics, and numerous researchers in academia and industry have shifted their attention to the relative and absolute quantification of glycoconjugates based on MS.

Several researchers have developed glycoprotein-specific enrichment combined with multiple-reaction monitoring (MRM) techniques for the absolute quantification of tissue inhibitor of metalloproteinase 1 (TIMPl) in the serum of colorectal cancer (CRC) patients. They used a systematic strategy coupling of phytohemagglutinin-L4 (L-PHA) lectin enrichment, stable isotope standards and capture by anti-peptide antibodies (SISCAPA) with MRM mass analysis to provide absolute quantification of TIMP1 at a molar (10-18) concentrations. This strategy provides a useful tool for the absolute quantitative analysis of specific abnormal glycoforms from human serum containing multiple protein isoforms.

Fig.1 Analytical workflow for the absolute quantification of N-glycosylated alpha fetoprotein with stable isotope-labeled N-glycopeptide by liquid chromatography-parallel reaction monitoring (LC-PRM). (Kim, 2020)

Our Services

CD BioGlyco provides a technical platform for absolute quantitative analysis of glycoproteins. We design and optimize the experimental process according to the specific requirements of customers. Our services include but are not limited to:

• Absolute quantification of glycans derivatized with reductive amination chemistry.
• Absolute quantification of glycans derivatized with reagents that introduce a cationic charge.
• Absolute quantification of N-glycosylation of glycoproteins using parallel reaction monitoring with stable isotope-labeled N-glycopeptide as an internal standard.
• Absolute quantification of neutral N-linked oligosaccharides by carboxymethyl trimethylammonium hydrazide modification and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

Applications

• Research on glycoprotein structure and function
• Study on glycan variability in proteins
• Discovery and validation of glycoprotein
• Systematic research on glycoproteomics

Advantages of Us

• Advanced and diverse technology platforms
• Highly trained research staffs
• High level of accuracy and consistency
• One-stop service

CD BioGlyco has developed absolute quantification strategies for glycoproteins based on advanced quantification techniques in proteomics and successfully applied them in scientific research. Our efficient work and attentive service attitude will facilitate our customers' research. If you have a demand for our services, please contact us for more information.

Reference

1. Kim, K.H.; et al. Absolute quantification of N-glycosylation of alpha-fetoprotein using parallel reaction monitoring with stable isotope-labeled N-glycopeptide as an internal standard. Analytical Chemistry. 2020, 92(18): 12588-12595.