Quantitative Monosaccharaide Analysis Service
High-level Monosaccharide Quantitative Analysis Service at Creative Biolabs
Abnormal glycosylation is related to many types of cancer. Cancer cells will gradually release proteins with abnormal glycosylation on their surface into body fluids. Therefore, the monosaccharide analysis in liquid-like molecules helps reveal changes in glycosylation, thereby discovering candidate biomarkers associated with disease. Creative Biolabs has a powerful Glycoprotein Analysis Platform and technical teams, and we provide high-quality monosaccharide quantitative analysis services to clients around the world. Quantitative analysis of monosaccharides in glycoproteins is of great significance for understanding the structure and functions of glycoproteins, ensuring quality control, disease diagnosis, and biomarker research.
Quantitative analysis of monosaccharides in glycoproteins typically requires the following steps:
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Release of monosaccharides
We use appropriate glycosidase (e.g. β-mannosidase, α-galactosidase, etc.) or PNGase F to hydrolyze glycoproteins. These enzymes cleave the sugar group's linkage to the protein, releasing the monosaccharides and protein moieties. In addition, under acidic conditions, sugar groups in glycoproteins are also released by hydrolysis, depending on the acidolysis conditions used.
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Derivatization of monosaccharides
To increase the detection sensitivity and selectivity of monosaccharides, we often need to derivatize monosaccharides, such as acetylation.
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Separation and detection of monosaccharides
We use a variety of techniques to separate various monosaccharides and conduct quantitative analysis of the separated monosaccharides. We have developed different analytical methods to efficiently and accurately determine the monosaccharide composition in glycoproteins. Commonly used detection methods include but are not limited to the following.
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We combine high-performance anion exchange chromatography with a pulsed amperometric detector (HPAEC-PAD), which is a simple and convenient method that analyzes the monosaccharide composition in glycoproteins sensitively and quickly without the need for pre-column or post-column solute derivatization.
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We use high-performance liquid chromatography with fluorescence detection (HPLC-FL) to analyze monosaccharides in glycoproteins. We select 2-aminobenzoic acid (2-AA) and 2-aminobenzamide (2-AB) as fluorescent labels, which are highly fluorescent and reactive, and the detection of monosaccharides is achieved by labeling carbohydrates with reducing ends before the column.
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We also couple HPLC with mass spectrometry to achieve monosaccharide analysis in glycoproteins. Commonly used ionization methods include electrospray ionization (ESI) or chemical ionization (CI). We use the mass spectrometry data collected by the mass spectrometer to confirm and quantify the monosaccharides in the sample.
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In addition, laser-induced fluorescence capillary electrophoresis (CE-LIF) technology also realizes monosaccharide analysis in glycoproteins. We use a laser to excite a fluorescent dye (usually a fluorescent probe that labels the monosaccharide) in the monosaccharide molecule, causing it to emit a fluorescent signal at a specific wavelength. The fluorescence signal is then captured and recorded by a detector to achieve an accurate analysis of monosaccharides in glycoproteins.
Quantitative analysis of monosaccharides in glycoproteins is a complex process. We combine a variety of technical means and analytical methods to ensure accurate determination of the content and composition of monosaccharides. In addition, we offer a variety of analytical services such as Sialic Acid Analysis, Sugar Nucleotide Analysis, and Glycosaminoglycan Analysis services.
Fig.1 Detailed steps for quantitative analysis of monosaccharides.
Publication
Technology: HPLC-FL
Book: Capillary Electrophoresis of Carbohydrates: From Monosaccharides to Complex Polysaccharides
Published: 2011
Results: The author chose 2-aminobenzoic acid as a fluorescent label, then directly derivatized monosaccharides with reducing ends through a reductive amination reaction, and then directly analyzed the monosaccharide components through HPLC-FL technology. The results showed that the chromatograms of recombinant IgG expressed in CHO cells were consistent with those of standard monosaccharides. This detection method can achieve highly accurate and reproducible monosaccharide analysis.
Fig.2 Monosaccharide analysis results by HPLC-FL.1
Advantages
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High sensitivity: We provide low-concentration monosaccharide detection services.
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High accuracy: We provide precise monosaccharide content analysis results.
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High reliability: Our analytical data is highly reproducible.
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Diversity: We provide monosaccharide analysis for various types of glycoproteins.
Application
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Food industry: Monosaccharide analysis is used to detect the content of glucose, fructose, and other monosaccharides in food, thereby guiding food processing and adjusting product taste.
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Biomedicine: Monosaccharides are components of cell surface sugar groups and are related to key biological processes such as cell-cell interactions and signal transduction. Monosaccharide analysis can understand the composition and changes of sugar groups on the cell surface, which helps study the mechanisms of disease.
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Environmental monitoring: Monosaccharide analysis is used to monitor microbial activities in water, soil, and other environments to provide data support for environmental pollution monitoring.
Creative Biolabs has extensive experience in monosaccharide analysis and can provide personalized monosaccharide analysis services according to client needs. In addition, we provide clients with comprehensive data analysis and result interpretation. Please do not hesitate to contact us if you would like to acquire detailed service.
Reference
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Mechref, Yehia. "Monosaccharide Compositional Analysis of Glycoproteins and Glycolipids: Utility in the diagnosis/prognosis of Diseases." Capillary Electrophoresis of Carbohydrates: From Monosaccharides to Complex Polysaccharides (2011): 237-267.
For Research Use Only.
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