Glycoprotein Quantitation Service

Glycoprotein quantitation involves measuring the levels of glycosylated proteins in biological samples. Glycosylation is a crucial post-translational modification where sugar molecules covalently attach to proteins, influencing their structure, function, stability, and intercellular interactions. Alterations in glycosylation patterns are strongly associated with various diseases, including cancer, diabetes, and autoimmune disorders. Quantitative analysis of glycoproteins aids in elucidating disease mechanisms, identifying potential biomarkers, and assessing therapeutic efficacy. Furthermore, glycoprotein quantification is essential in drug development and biopharmaceutical quality control, ensuring the safety and effectiveness of therapeutic agents. However, the presence of multiple glycosylation forms on a single protein, the diversity and complexity of sugar moieties, and the difficulty in detecting low-abundance glycoproteins pose significant challenges. Current analytical techniques often struggle with resolving and quantifying multiple glycosylation sites, necessitating the development of more advanced and sensitive methods for accurate glycoprotein quantification.

 

Analysis Workflow

 



 

Service Advantages

1. Advanced Instrumentation Ensuring High-Sensitivity Detection

MtoZ Biolabs is equipped with state-of-the-art mass spectrometers and liquid chromatography systems, enabling precise detection of glycoproteins at extremely low concentrations, with detection limits down to the nanogram level. These advanced instruments ensure high data reliability and accuracy.

 

2. Innovative Methods Enhancing Detection Efficiency

Utilizing optimized glycoprotein extraction and purification techniques, we preserve the natural structure and function of glycoproteins to the greatest extent. The refined analytical processes effectively reduce signal loss and background interference, significantly improving detection sensitivity and accuracy of results.

 

3. Professional Technical Team Offering Customized Services

MtoZ Biolabs boasts an experienced professional team with deep expertise in glycoproteomics and mass spectrometry analysis. Our experts provide personalized technical support and tailored solutions to ensure the successful advancement of your research projects and the attainment of anticipated objectives.

 

Sample Submission Suggestions

1. Sample Type: We accept various types of glycoprotein samples, including single glycoproteins, glycopeptides, glycosylated proteins, and complex glycosylation-modified proteins. Samples can be naturally derived glycoproteins from cells, tissues, or fluids, or recombinant glycoproteins expressed in vitro.

 

2. Sample Storage: Glycoproteins should be stored at low temperatures, ideally at -80°C. Avoid repeated freeze-thaw cycles to prevent degradation and inactivation of the glycoproteins. Samples should be stored in appropriate buffers that maintain their stability, such as PBS (phosphate-buffered saline) or Tris-HCl buffers, with a pH range typically between 7.0-8.0.

 

3. Sample Purity: To ensure accurate quantification, it is recommended to provide high-purity glycoprotein samples, free from contaminants such as salts, lipids, or other small molecules that may interfere with the analysis. If contaminants are present, purification steps should be performed prior to submission.

 

4. Sample Submission: Samples should be submitted in appropriate containers (such as lyophilized powder or small volume solutions) with clear labeling and detailed information, including sample type, concentration, and storage conditions. Please also provide background information on the sample to help us optimize the analysis plan.

 

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

 

Applications

1. Integrated GlycoProteome  Analyzer (I-GPA) for Automated  Identification and Quantitation of  Site-Specific N-Glycosylation

 



Park, GW, et al. Sci Rep. 2016.

 

2. Identification and Quantification of N-Linked Glycoproteins Using Hydrazide Chemistry, Stable Isotope Labeling and Mass Spectrometry

 

 



Zhang, H, et al. Nat Biotechnol. 2003.

 

FAQ

1. How can glycoproteins be efficiently extracted and purified to avoid protein loss or alterations in glycosylation sites during sample processing, thereby ensuring the accuracy of quantitative results?

To achieve efficient extraction and purification of glycoproteins while preventing protein loss and changes in glycosylation sites, it is advisable to use buffers containing non-ionic or zwitterionic surfactants to minimize structural alterations to glycoproteins. Harsh chemical or physical treatments should be avoided. Experiments should be conducted at low temperatures (4 °C) and appropriate pH conditions, with the addition of protease and glycosidase inhibitors to prevent degradation by endogenous enzymes. Reducing processing time can further decrease the likelihood of glycosylation site modifications due to external factors. Employing lectin affinity chromatography allows for selective enrichment of glycoproteins, reducing interference from non-target proteins and enhancing purification efficiency. Over-processing should be minimized by reducing steps such as centrifugation and transfer to lower the risk of sample loss.

 

2. Given the diversity of glycosylation forms in glycoproteins, how can reliable standard curves and calibration methods be established to accurately reflect the impact of different glycoforms and glycosylation levels on quantitative results?

To accurately account for the effects of glycosylation diversity on quantification, glycosylated standards should be utilized. Selecting standard glycoproteins with known glycoforms and degrees of glycosylation allows for the construction of specific standard curves. Correction factors can be calculated based on differences in the ionization efficiency of various glycoforms to adjust quantitative results accordingly. It is important to conduct methodological validation by evaluating the linear range, sensitivity, and reproducibility to ensure the reliability of the quantitative methods. When processing data, integrating mass spectrometry with glycomics databases facilitates comprehensive analysis of glycosylation patterns, thereby improving the identification and quantification of different glycoforms.

 

Deliverables

1. Comprehensive Experimental Details

2. Materials, Instruments, and Methods

3. Relevant Liquid Chromatography and Mass Spectrometry Parameters

4. The Detailed Information of Glycoprotein Quantitation Analysis

5. Mass Spectrometry Image

6. Raw Data