Absolute Quantification of Glycoprotein
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 Workflow for absolute quantification of proteins. (Rzagalinski, et al., 2022)
Absolute Quantification of Glycoproteins: Unlocking the Molecular Code of Complex Biological Samples
CD BioGlyco provides a comprehensive range of absolute glycoprotein quantification services, utilizing industry-leading technology to accurately determine the levels of specific glycoproteins in your samples, enabling you to study their biological functions in depth.
- Mass Spectrometry (MS)-based methods
Using advanced selective reaction monitoring (SRM/MRM) or data-independent acquisition techniques in combination with stable isotope labeling strategies, we provide you with high sensitivity, specificity and accuracy for absolute quantification of glycoproteins. We are particularly good at the precise determination of low-abundance glycoproteins in complex samples and the analysis of glycosylated isomers.
- Lectin-based approach
Through professional lectin affinity chromatography, combined with subsequent quantitative analysis, we provide clients with glycoprotein enrichment and absolute quantification services for specific glycan modifications and help clients accurately analyze the impact of glycosylation on protein function.
- Other methods
- 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.
Workflow
Applications
- Our absolute quantitative glycoprotein analysis service can be used for characterization, quality control, and batch consistency monitoring of biologics.
- Our absolute quantitative glycoprotein analysis service can be used in the drug development process to analyze glycoproteins for structure-function relationship studies and to guide drug design and optimization.
- Our absolute quantitative glycoprotein analysis service can be used for mechanistic studies of glycosylation modifications, exploring the role of glycoproteins in cell signaling, immune response, and other processes.
Advantages of Us
- We have a professional team of senior scientists and technicians who have rich experience and deep technical skills in the field of glycoprotein analysis, and can provide you with high-quality services.
- Our company has established a perfect quality control system, from sample reception, experimental operation, to data analysis, each link is strictly under quality control to ensure the reliability of the experimental results.
- Our team of researchers and data analysts has years of successful experience in helping clients complete projects quickly.
Publication Data
Journal: Journal of Proteome Research
Published: 2022
IF: 3.8
Results: This document describes an innovative method called FastCAT for accelerated absolute quantitative analysis of proteins. By utilizing multiple short, stable isotope-labeled chimeric protein standards (CPs), the researchers achieved direct quantification of target proteins in biological samples without the need for purified CPs. This approach significantly improves the accuracy, precision, and dynamic range of quantification and has successfully quantified a variety of neuromarkers in human cerebrospinal fluid (CSF).
Fig.2 The truncation pattern of chimeric proteins. (Rzagalinski, et al., 2022)
Frequently Asked Questions
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.
Associated Services
Reference
- Rzagalinski, I.; et al. FastCAT accelerates absolute quantification of proteins using multiple short nonpurified chimeric standards. Journal of Proteome Research. 2022, 21(6): 1408-1417. (Open Access)
