Glycomic Profiling Service

What Is Glycomic Profiling

Glycomics, at its core, involves the collection, analysis, and exploitation of glycan-related biological data on a glycome-wide scale. The profound impact of glycosylation on lipids and proteins has elevated glycomics to a pivotal realm within the life sciences. Global glycomic profiling, which encompasses the comprehensive examination of all glycans, serves as a valuable complement to genomics and proteomics. When applied at the cellular or organismal level, glycomics provides a holistic perspective on the glycome—the complete glycosylation patterns of glycoproteins, lipids, and other biomolecules. This glycomic profiling approach proves invaluable for comparing distinct glyco-engineered host strains for therapeutic glycoprotein production or assessing various stages of cancer, offering potential insights into novel biomarkers.

The goal of glycomic profiling is to identify, quantify, and characterize diverse glycan types, structures, abundance, modifications, and glycan-binding proteins within a given sample. This comprehensive analysis encompasses N-glycans (attached to asparagine residues of proteins), O-glycans (attached to serine or threonine residues of proteins), and glycolipids (glycans attached to lipids).

Glycomic profiling techniques typically include a combination of analytical methods such as mass spectrometry, liquid chromatography, and various glycan-specific binding assays (lectin arrays, antibodies, etc.).

What Can We Do

Creative Proteomics offers comprehensive glycoproteomics services for intact glycopeptides, enabling the simultaneous analysis of N-glycan and O-glycan glycosylation sites, glycan types, glycopeptides, and glycoproteins through efficient enrichment techniques and authoritative analytical software. As a result, Creative Proteomics has established a comprehensive glycoproteomics service encompassing N-glycosylation modification, O-glycosylation modification, and the entire glycoproteome.

Glycomic Profiling Service by MALDI TOF MS

This service is for the identification of an entire set of N-glycans expressed by plasma/serum, cell, tissue or organism. All glycans attached to the proteins will release by enzymatic digestion, and then separated by hydrophilic chromatography and finally quantitatively profiled with MALDI-TOF MS system. Steps are briefly showed as below:

Sample preparation
glycoprotein and/or glycolipid fractions isolation
N-, O- and/or glycosphingolipid glycans release
Permethylation of glycans
Analysis of native and permethylated glycans by MALDI-TOF MS
Glycan structures confirmation by MS/MS fragmentation if required
Statistical Analysis for the potential biomarker glycans.

Delivery

1. Information of the native and permethylated glycans from your samples;

2. Structure of the glycans from the sample.

Advantages of Our Glycomic Profiling Service

Comprehensive Glycome Analysis: We employ rapid and efficient chemical derivatization labeling along with glycopeptide enrichment methods to provide in-depth profiling of the glycome.

Complete Glycopeptide Analysis: Our approach includes comprehensive glycopeptide analysis, allowing us to simultaneously obtain information on glycan sites, glycan types, and paired qualitative and quantitative data.

Non-depletive, Minimal Sample Requirement: We utilize non-depletive methods that require minimal original sample volume, resulting in relatively low experimental error.

High-Throughput, High Sensitivity Mass Spectrometry: Our service leverages high-throughput and high-sensitivity mass spectrometry analysis, leading to shorter service turnaround times.

Applications of Glycomic Profiling

Glycomic profiling finds significant applications across various domains, encompassing:

Disease Biomarker Discovery: Alterations in glycan profiles have been linked to a spectrum of diseases, including cancer, autoimmune conditions, and infectious ailments. Glycomic profiling serves as a valuable tool for identifying potential biomarkers facilitating disease diagnosis, prognosis, and monitoring.

Glycoprotein Analysis: Numerous proteins, particularly those found on cell surfaces and antibodies, undergo glycosylation. Glycomic profiling can be employed to scrutinize the glycosylation patterns of specific glycoproteins, thereby shedding light on their functionalities and contributions to disease mechanisms.

Vaccine Development: Glycan analysis plays a pivotal role in comprehending the glycosylation patterns of pathogens, a crucial asset in the development of vaccines and therapeutic agents.

Biopharmaceuticals: Profiling the glycosylation of therapeutic proteins, such as monoclonal antibodies, assumes paramount importance in ensuring their safety and efficacy.

How to place an order:

*If your organization requires signing of a confidentiality agreement, please contact us by email

As one of the leading companies in the omics field with over years of experience in omics study, Creative Proteomics provides glycomics analysis service for our customers. Contact us to discuss your project!

Comprehensive glycoprofiling of oral tumors associates N-glycosylation with lymph node metastasis and patient survival

Journal: Molecular and Cellular Proteomics
Published: 2023
Impact Factor: 7.4
Research Material: Primary OSCC Tumors
Techniques: Glycomics, Glycoproteomics

Results

01 N-Glycomics Analysis of OSCC Tissues

This study aimed to compare the N-glycan alterations between OSCC tissues with (N+) and without (N0) lymph node metastasis. The identified N-glycans in OSCC tissues predominantly comprised complexes, oligomannosides, and paucimannosides (Figure 1A). In comparison to N0 tissues, six N-glycans were found to be differentially expressed in N+ tissues (Figure 1B), and each of these differentially expressed N-glycans could stratify N0 and N+ patients through logistic regression and random forest models (Figure 1C).

Figure 1: Glycomics Analysis (Carnielli CM., Molecular and Cellular Proteomics, 2023)

02 N-Glycoproteomics Analysis of OSCC Tissues

In this investigation, N-glycoproteomics was harnessed to discern variations in the glycoprotein composition within tumor tissues of N+ and N0 patients. The analysis unveiled a broad spectrum in the distribution of N-glycans and N-glycoproteins in OSCC tissues. Predominantly, the N-glycans associated with N-glycopeptides were characterized by complex structures or oligomannosides (depicted in Figure 2A-B). Notably, the glycoproteomic data indicated no substantial disparities in N-glycan distribution between N+ and N0 patients, wherein the majority of N-glycosylation sites exhibited representation by more than one N-glycan component. Conversely, most glycoproteins were observed to feature a solitary N-glycosylation site (Figure 2C-D). Furthermore, researchers detected 79 differentially expressed N-glycopeptides within the two patient cohorts, all of which exhibited the potential for stratification (Figure 2E-F).

Figure 2: Glycoproteomics Analysis (Carnielli CM., Molecular and Cellular Proteomics, 2023)

03 Clustering Analysis of OSSC Tissues

The primary objective of this investigation was to conduct a comprehensive analysis of N-glycosylation patterns within primary OSSC tumor tissues. Utilizing clustering analysis, we identified two predominant tumor clusters, denoted as T-C1 and T-C2, as well as two distinct N-glycan clusters, referred to as NG-C1 and NG-C2. Within NG-C1, there were discernible alterations in the distribution of various N-glycan classes between T-C1 and T-C2, notably affecting the relative abundance of paucimannosides and complex-type N-glycans (see Figure 3A).

In parallel, our analysis of N-glycoproteomics similarly revealed two prominent tumor clusters (T-C1 and T-C2) and two N-glycopeptide clusters (IG-C1 and IG-C2). Within IG-C1, we observed differential expression patterns in highly truncated, oligomannosides, and complex-type N-glycans between T-C1 and T-C2 (as depicted in Figure 3B). Conversely, within IG-C2, the distinction between the two tumor groups was primarily observed in the context of highly truncated N-glycans.

Figure 3: Clustering Analysis (Carnielli CM., Molecular and Cellular Proteomics, 2023)

04 Relationship Between N-Glycoproteome Composition, Clinical Features, and Biological Processes

The primary aim of this investigation was to assess potential associations between differentially expressed N-glycans and N-glycopeptides and various clinical characteristics. Utilizing clustering analysis of N-glycans and N-glycopeptides, notable distinctions were observed concerning vascular invasion, lymph node status, and tumor size, as depicted in Figure 4A-D.

To elucidate potential connections between N-glycosylation patterns and specific biological processes, cellular components, and molecular functions, the study employed Gene Ontology (GO) functional annotation. This comprehensive analysis revealed that IG-C1 exhibited enrichment in functions related to cell-matrix adhesion and the collagen-containing extracellular matrix. Conversely, IG-C2 demonstrated enrichment in functions associated with fibroblast activation, proteinase binding, and other pertinent biological processes.

Figure 4: Clustering Analysis and GO Functional Annotation (Carnielli CM., Molecular and Cellular Proteomics, 2023)

05 The Association of N-Glycans and N-Glycopeptides with Clinical Outcomes

The primary aim of this study was to elucidate the relationship between variations in N-glycans and N-glycopeptides and the survival outcomes of patients. Within OSSC tumor tissues, discernible alterations were observed at both the protein and glycan levels, as depicted in Figure 5A-B.

Survival analysis using Kaplan-Meier plots provided compelling evidence of an association between elevated levels of Glycan 40a and Glycan 46a and diminished patient survival rates, as illustrated in Figure 5C. Additionally, three distinctively expressed N-glycopeptides exhibited correlations with patient survival when comparing N0 and N+ patient groups, as presented in Figure 5D.

Figure 5: The Association of N-Glycans and N-Glycopeptides with Patient Survival Rates (Carnielli CM., Molecular and Cellular Proteomics, 2023)

Conclusion

This investigation, employing N-glycomics and N-glycoproteomics analyses, has illuminated the relationships between protein N-glycosylation in OSSC tumor tissues and critical clinical parameters, as well as patient survival rates. It has elucidated the intricate and dynamic alterations within OSSC tissues in the presence or absence of lymph node metastasis. Consequently, these findings offer valuable insights into the fundamental mechanisms of the disease and hold promise for identifying potential prognostic glyco-markers in the context of OSCC exploration.