3-O-Sulfation Analysis Service

Q: How does the LC-MS/MS method improve the detection of low-abundance 3-O-sulfation?

Our LC-MS/MS method is optimized with 13C-labeled standards to precisely quantify low-abundance 3-O-sulfation in complex samples, improving sensitivity and accuracy.

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Specialized 3-O-Sulfation Analysis Service at Creative Biolabs

Creative Biolabs has built a mature Polysaccharide Analysis platform, and we provide reliable Heparan Sulfate (HS), 3-O-sulfation analysis, and Fiber Analysis services.

Sample pretreatment

We perform pre-treatment steps such as desalting and centrifugation on the samples to ensure that the samples have sufficient purity and quality.

Degradation reaction

We use appropriate enzymes or chemical reagents to degrade the sample into smaller fragments in order to detect 3-O-sulfation modification sites.

Separation and analysis

We separate different fragments in the sample using techniques such as high-performance liquid chromatography (HPLC) or ion exchange chromatography (IEC). Moreover, mass spectrometry (MS) and nuclear magnetic resonance (NMR) technology are combined to conduct qualitative and quantitative analysis of 3-O-sulfation in different fragments by detecting specific ions or ion fragments. In addition, we set up controlled experiments to verify the existence of 3-O-sulfation through multiple experiments and repeated testing.

At present, we have developed a fast LC-MS/MS method to quantitatively determine low-abundance 3-O-sulfation in HS through ¹³C-labeled 8-mer and 10-mer standards. Since these standards are similar to the 3-O-sulfation structural motif in enzymatically digested HS, this strategy is suitable for analyzing different samples.

Professional 3-O-sulfation analysis service. (Creative Biolabs Original)

Published data

As a new analytical technique for glycomics, MS is highly sensitive and identifies the position of sugar termini, distinguishes the connection mode, and determines the position of substituents such as sulfate. In this study, the authors used zebrafish as a model to study its glycobiological characteristics during development. The authors selected three organs (i.e., brain, intestine, and ovary) of adult zebrafish and extracted proteins from these organs, then digested them with chymotrypsin and trypsin, and then released N-sugars by PNGase F, while O-sugars were released by reductive elimination. Subsequently, the authors used LC-MS/MS technology to analyze the sulfated N- and O-glycans of interest and their glycosylation sites in zebrafish. The results demonstrated the presence of sulfated sugar sites in zebrafish, mainly sulfated N-acetylgalactosaminyl-β1,4-N-acetylglucosamine (LacdiNAc) and 3-O-sulfated galactose (Gal).

Fig.1 Sulfated N-glycan analysis results of zebrafish. Fig.1 Sulfated N-glycan profile of adult zebrafish.¹

Advantages

Professional teams: We have an experienced team of scientific researchers and experts who perform 3-O-sulfation analysis accurately and efficiently.
Diversified analysis: We use a variety of advanced instruments to provide comprehensive 3-O-sulfation analysis services, including structural analysis, quantitative analysis, and other aspects.
Efficient service: We actively respond to client needs, and provide rapid 3-O-sulfation analysis services. Moreover, we provide clear and comprehensive data interpretation and reporting.

Application

Carbohydrate research: Analyzing the position and degree of 3-O-sulfation in the carbohydrate structure helps to study the impact of carbohydrate sulfation modifications (such as heparan sulfate) on cell signaling, cell-cell interactions, and disease development.
Biological drug development: Analysis of 3-O-sulfation modifications in biological drugs (such as protein drugs) can help evaluate the stability, activity, and immunogenicity of drugs, thereby guiding drug design, optimization, and quality control.
Structural biology: 3-O-sulfation analysis helps reveal the location and structure of 3-O-sulfation modifications in proteins, sugars, or other biological macromolecules, thereby gaining an in-depth understanding of the functions and interactions of biological molecules.

Creative Biolabs has professional analytical technical teams and a variety of advanced instruments. We have a quality control system and standardized analysis processes to provide clients with accurate and reliable 3-O-sulfation analysis services. Please feel free to contact us if you would like to inquire about specific analysis content.

FAQ

Q1: What types of samples can be analyzed for 3-O-sulfation?

A1: 3-O-sulfation analysis mainly studies carbohydrates and polysaccharides in biological samples. These samples include but are not limited to, glycans in glycoproteins, glycosaminoglycans in proteoglycans (such as heparin and chondroitin sulfate), and free oligosaccharides and polysaccharides. 3-O-sulfation analysis can provide insights into the structural characteristics of these compounds and their biological functions, such as regulation of cell signaling, protein binding, and metabolic processes in the body. This analysis is particularly important for studying biomacromolecules, especially those involving complex glycan modifications.

Q2: What methods do you use for sample pretreatment in 3-O-sulfation analysis?

A2: Sample pretreatment in 3-O-sulfation analysis usually involves removing impurities in the sample that may interfere with the analysis, which can be achieved in a variety of ways, such as purification by solid phase extraction (SPE). In addition, the sample may need to be degraded or enzymatically treated to release the target compounds for subsequent detection and analysis. Sample concentration and drying steps are also often applied to improve the sensitivity and accuracy of the analysis. The combination of these methods can ensure that the sample reaches the optimal purity and condition before entering the analytical instrument.

Q3: How does the LC-MS/MS method improve the detection of low-abundance 3-O-sulfation?

A3: Our LC-MS/MS method is optimized with ¹³C-labeled standards to precisely quantify low-abundance 3-O-sulfation in complex samples, improving sensitivity and accuracy.

Customer Review

Seamless And Efficient Analysis Service
“Creative Biolabs provided a seamless and efficient service. Their LC-MS/MS method for detecting low-abundance 3-O-sulfation was particularly impressive, allowing us to gain insights into complex structures quickly, which greatly benefited our research.”

Effective Communication And Professional Analysis
“Effective communication and rapid service are reasons I would recommend Creative Biolabs. Creative Biolabs handled our project with utmost professionalism. They kept us informed at every stage and promptly delivered comprehensive analysis reports, which was crucial for our decision-making process in further experiments.”

Reference

Tseng, Huan-Chuan, et al. "Discovery Sulfoglycomics and Identification of the Characteristic Fragment Ions for High-Sensitivity Precise Mapping of Adult Zebrafish Brain–Specific Glycotopes." Frontiers in Molecular Biosciences 8 (2021): 771447. Distributed under Open Access license CC BY 4.0, without modification.

For Research Use Only.