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Liquid Chromatography-Electrospray Ionization-Mass Spectrometry (LC-ESI-MS)

With years of experience in the field of glycoprotein analysis, Creative Biolabs is able to offer a wide range of glycoprotein analysis services for customers all over the world. LC-ESI-MS is a sensitive analytical technique that has been widely used for glycoform profiling at the level of glycopeptides. Based on this technique, we can offer the most comprehensive information of protein glycosylation site(s) and occupancy of glycopeptide/glycoprotein.

Glycoproteins Analysis Based on LC-ESI-MS

LC-ESI-MS is a hyphenated mass spectrometry technique. This method combines the resolution of high-performance liquid chromatography (HPLC) separation with the high mass accuracy of the mass spectrometer. Therefore, it is used for the determination of intact protein mass or for in-depth analysis of protein or peptide mixtures. Through the coupling with HPLC, the detected characteristics can be quantified by ultraviolet light or MS signal intensity. Moreover, this method has been used for glycoform profiling of more complex therapeutic proteins, such as erythropoietin with one O- and 3 N-glycosylation sites. In the analysis of N-glycoproteins, N-glycoproteins are enriched using hydrazide chemistry and lectin capture to remove nonglycoproteins, following digested by trypsin and PNGase F treatment. Then the generated glycopeptides or deglycosylation peptides or nonglycosylation peptides are identified and quantified by LC-ESI-MS/MS. The protein glycosylation site(s) and occupancy of glycopeptide can be achieved by this method. Furthermore, ESI-MS is a very important tool for glycoform profiling at the glycopeptide level. Importantly, prior to MS analysis, it is usually required to enrich glycopeptides by HPLC to overcome ion suppression of glycopeptide signals by regular peptide signals. Recently, LC-ESI-CID-MS/MS has been widely applied to glycan sequencing, and it shows improved speed and sensitivity.

LC-ESI-MS Analysis Services

Creative Biolabs has focused on the glycoprotein analysis for many years. Based on our advanced techniques, we are capable to offer a full range of glycoprotein analysis services for clients all over the world. LC-ESI-MS is our innovative and excellent platform that has been widely used for Glycomic Profiling. Based on LC-ESI-MS technology, we are able to offer the best glycomic profiling service. Besides, our experienced experts also provide a wide range of supports, including analytical method development, method validation, method transfer, and stability testing.

Features of Our Services

  • Excellent technical knowledge
  • Long-standing experience
  • High quality service
  • Sensitive and high-flux technique platforms
  • Comprehensive technical guidance
  • Accurate and comprehensive analysis data

Equipped with the versatile technology platform and experienced expert team, Creative Biolabs is confident in providing the most comprehensive glycoprotein analysis services for global clients at a competitive price. Besides, we offer custom glycoprotein analysis services based on customers’ requirements. If you are interested in our LC-ESI-MS analysis services, please feel free to contact us for more details.

Published data

Mild cognitive impairment (MCI) may increase the risk of developing Alzheimer's disease (AD). Therefore, accurately predicting clinical changes in MCI patients is of great significance for early diagnosis of AD and monitoring disease progression. Scientists have found that changes in serum N-glycan expression may play a vital role in the pathophysiology of neurodegenerative diseases and can serve as a source of potential biomarkers for evaluating MCI diseases. In this study, the authors collected serum samples from MCI patients and healthy subjects and removed high-abundance proteins from the samples. Then the authors extracted N-glycans from serum samples and released, purified, reduced, and fully methylated N-glycans. Subsequently, the fully methylated N-glycans in different serum samples and the isomerized glycome of serum glycoproteins were analyzed by sensitive LC-MS/MS glycomics to better study low to medium-abundant proteins in serum samples. Multiple reaction monitoring (MRM) mass spectrometry analysis resulted showed that the authors found five isomerized N-glycans in serum samples, which were expected to be used as potential biomarkers to distinguish healthy people from MCI patients. The identified N-glycans may play important roles in neurodegenerative diseases and have the potential to distinguish disease states.

Fig.1 LC-MS/MS glycomic analysis process.Fig.1 LC-MS/MS glycomic analysis steps in serum samples.1

FAQs

Q1: How is glycan sequencing performed with LC-ESI-MS and what are the advantages?

A1: LC-ESI-MS is a powerful tool for glycan sequencing. In this process, glycopeptides are first separated by LC and then ionized using ESI. The ionized glycopeptides are then subjected to collision-induced dissociation (CID), which causes them to fragment predictably. Subsequently, the fragment ions resulting from the fragmentation are analyzed in the MS stage, allowing detailed sequencing of the glycan structure. The main advantages of this method are its enhanced sensitivity and speed, which enables the detection of trace glycan species and provides fast and detailed glycan structural information.

Q2: What information can be obtained about glycoproteins using LC-ESI-MS?

A2: Various detailed information about glycoproteins can be obtained using LC-ESI-MS, including:

  • Identification of glycosylation sites (N-linked and O-linked).
  • Characterization of the glycan structures attached to these sites.
  • Quantification of the occupancy of each glycosylation site.
  • Analyze different glycoforms of proteins.
  • Analyze changes in glycosylation patterns under different biological conditions or disease states.

Q3: Can you customize LC-ESI-MS analysis services to meet specific research needs or industrial applications?

A3: Yes, we provide customized LC-ESI-MS analysis services to meet specific research needs or industrial applications. Our team works closely with clients to understand their unique needs and develop custom methods to address specific research questions or challenges. Customization options include specific sample preparation protocols, targeted glycopeptide enrichment techniques, specialized LC and MS setups, and customized data analysis workflows. We also offer flexible project designs, ranging from small feasibility studies to large comprehensive glycoprotein profiling projects. We encourage clients to discuss their specific needs with our experts to develop a customized plan that ensures optimal results.

Customer Review

Detailed Glycosylation Site Analysis
"Creative Biolabs performed an extremely detailed analysis of the glycosylation sites of our glycoprotein samples. Their LC-ESI-MS platform provided unparalleled sensitivity, allowing us to pinpoint previously undetectable glycosylation sites. This information was critical to our studies of complex therapeutic proteins."

Highly Sensitive And Specific Analysis
"I was very pleased with the sensitivity and specificity of Creative Biolabs' LC-ESI-MS platform. The method was particularly effective in overcoming ion suppression issues, ensuring accurate detection of glycopeptide signals even in complex mixtures. I highly recommend their glycopeptide analysis services."

References

  1. Reyes, Cristian D. Gutierrez, et al. "LC-MS/MS isomeric profiling of N-glycans derived from low-abundant serum glycoproteins in mild cognitive impairment patients." Biomolecules 12.11 (2022): 1657. Distributed under Open Access license CC BY 4.0, without modification.
For Research Use Only.

Related Services:

  1. Lectin Microarray
  2. MS
  3. MALDI-TOF MS
  4. HPLC
  5. HPAEC-PAD
  6. RP-HPLC
  7. UHPLC/FLD/Q-TOF
  8. FTIR
  9. NMR
  10. SPRi
  11. TLC
  12. Flow Cytometry
  13. GC-MS
  14. Imaging Mass Spectrometry
  15. Glycobiology Microarray
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