Native N-glycome profiling Service

N-glycome profiling involves the qualitative and quantitative determination of N-glycan structures on proteins without undergoing chemical derivatization. N-glycan chains are complex glycosylation modifications formed by the covalent attachment of asparagine (Asn) residues to proteins. These modifications are widely present on cell surfaces and in secreted proteins, influencing protein stability, function, and protein-protein interactions. Traditional N-glycan analysis often requires derivatization of glycan chains to enhance detection signals or improve separation efficiency. However, this process can introduce structural alterations, increasing the complexity and duration of sample preparation. Native mass spectrometry techniques allow for the direct analysis of glycan chains without derivatization, thereby preserving their original structural information, simplifying sample preparation, reducing sample loss, and enhancing both analysis efficiency and accuracy. This approach is particularly suitable for high-throughput and high-sensitivity N-glycome profiling analysis, effectively addressing challenges related to glycosylation heterogeneity and complex glycan structures. Consequently, native MS ensures efficient and accurate detection of glycoproteins in complex biological samples.

 

Service at MtoZ Biolabs

MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider, provides advanced proteomics, metabolomics, and biopharmaceutical analysis services to researchers in biochemistry, biotechnology, and biopharmaceutical fields. We have established a powerful and professional native N-glycome profiling platform, which includes Thermo Fisher Q Exactive HF and Obitrap Fusion Lumos mass analyzer system, coupled with Nano-LC system. Our Native N-glycome profiling service is designed to provide high-quality glycome analysis, particularly focused on N-glycosylation modifications on proteins. By utilizing advanced mass spectrometry techniques, we are able to precisely analyze the glycan structures, composition, and distribution characteristics in samples without disrupting their native conformation. We are committed to delivering accurate, reliable, and comprehensive glycome data to support your research in glycosylation modification analysis. If you have related needs, feel free to contact us for a personalized analysis plan.

 

Analysis Workflow

 

 

Service Advantages

1. High Sensitivity for Low-Abundance Samples 

MtoZ Biolabs' native N-glycome profiling service enables the precise characterization of glycoproteins in single-cell and nanogram-level blood samples, with the ability to detect glycosylation even at very low concentrations. This provides an effective approach for studying rare sample types or trace sample where traditional methods are insufficient.

 

2. Superior Matrix Compatibility for Minimizing Interference

Our technology is optimized to handle complex biological matrices, such as blood and tissue, effectively mitigating matrix-related interferences to ensure the accurate identification and analysis of glycoproteins. This makes it particularly advantageous for cases where other methods are hindered by matrix effects.

 

3. Tailored Analytical Protocols to Ensure Accuracy and Consistency

MtoZ Biolabs has developed specialized workflows, optimized for different sample types and conditions, ensuring the robustness and accuracy of results. This approach not only addresses the challenges of low-concentration detection but also offers customized solutions to meet diverse research requirements.

 

Sample Submission Suggestions

1. Sample Type: The provided sample should be non-denatured glycoproteins. It can be purified glycoproteins or glycoproteins extracted from cell or tissue samples. The sample should be clean, free from contaminants, and preferably retain its native conformation.

 

2. Sample Storage: Samples should not be subjected to high temperatures, acidic or basic conditions, or strong oxidizing agents. To preserve glycan structures, samples should be stored at -80°C or 4°C. If cryopreserved with liquid nitrogen, ensure the sample remains frozen during transport.

 

3. Sample Buffer: Use buffers that do not contain detergents, metal ion chelators, or other substances that could interfere with glycan profiling. Commonly used buffers include PBS (phosphate-buffered saline) or Tris-HCl buffer.

 

4. Sample Preparation: If the sample is from cell or tissue extracts, it is recommended to purify the glycoproteins and remove impurities before analysis to improve sample purity. For complex glycoproteins, purified proteins are preferred for analysis.

 

If you need more detailed information or specific analytical methods, feel free to ask!

 

Sample Results

1. Native N-glycome Profiling of Single Cells and Ng-Level Blood Isolates Using Label-Free Capillary Electrophoresis-Mass Spectrometry¹

 



 

FAQ

1. How can glycoproteins be efficiently extracted and purified from single cells or nanogram-level blood samples while preserving their native conformation and activity?

To achieve efficient extraction and purification of glycoproteins from micro-scale samples while maintaining their native conformation and activity, it is crucial to optimize lysis methods. Employing gentle lysis buffers containing non-ionic surfactants can minimize protein degradation. Additionally, combining affinity purification techniques such as lectin affinity chromatography can effectively enrich target glycoproteins. All experimental procedures should be performed at 4°C to prevent protein degradation and denaturation.

 

2. For samples with extremely low concentrations, which high-sensitivity analytical techniques can accurately detect and quantify N-glycans, and how can signal loss and background interference be minimized during the analysis process?

To accurately detect and quantify N-glycans in low-concentration samples, high-sensitivity mass spectrometry techniques like nanoLC-MS/MS with high-resolution instruments should be utilized to enhance detection sensitivity. Derivatization or labeling of N-glycans can further amplify mass spectrometric signals. Furthermore, implementing solid-phase extraction or lectin-based enrichment methods can increase the relative abundance of the target analytes, thereby reducing signal loss and background interference.

 

Deliverables

1. Comprehensive Experimental Details

2. Materials, Instruments, and Methods

3. Relevant Liquid Chromatography and Mass Spectrometry Parameters

4. The Detailed Information of N-glycome Profiling Analysis

5. Mass Spectrometry Image

6. Raw Data

 

Reference

1. Marie AL, Gao Y, Ivanov AR. Native N-glycome profiling of single cells and ng-level blood isolates using label-free capillary electrophoresis-mass spectrometry. Nat Commun. 2024 May 8;15(1):3847.