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How Glycan Analysis Supports Biopharmaceutical Development

Posted on by Asparia Glycomics

Glycosylation plays a critical role in the efficacy, safety, and stability of biopharmaceuticals. As the biopharma industry grows, glycan analysis and profiling have become essential for ensuring product quality and regulatory compliance. At Asparia Glycomics, we specialize in advanced glycan analysis services, providing precise and reliable data to support biopharmaceutical development.

In this article, we explore how glycan analysis enhances biopharmaceutical development, its impact on drug performance, and why Asparia Glycomics is your trusted partner for glycosylation analysis and profiling.

The Importance of Glycan Analysis in Biopharmaceuticals

Glycans influence key properties of therapeutic proteins, including:

  • Pharmacokinetics –The glycan structures attached to biopharmaceuticals play a crucial role in determining their serum half-life and clearance rates. For instance, terminal sialic acid residues on N-glycans help shield proteins from rapid uptake by the liver’s asialoglycoprotein receptor (ASGPR), thereby extending circulation time. This is particularly important for drugs like erythropoietin (EPO) and therapeutic antibodies, where prolonged activity enhances efficacy. Conversely, high-mannose glycans can accelerate clearance via mannose receptors on macrophages, making glycan profiling essential for optimizing drug formulations. Another example of the stabilization effect of glycosylation is found in Fc-glycan modifications in monoclonal antibodies. It influences their interaction with neonatal Fc receptors (FcRn), which recycle antibodies and prevent lysosomal degradation. Hence, by fine-tuning glycosylation, biopharmaceutical developers can enhance drug bioavailability and reduce dosing frequency—key factors in patient compliance and cost-effectiveness.
  • Immunogenicity – Certain glycan structures, such as α-galactose (α-Gal) and N-glycolylneuraminic acid (Neu5Gc), are foreign to humans and can provoke immune reactions, leading to anti-drug antibodies (ADAs) that neutralize therapeutic effects. This is especially critical for recombinant proteins derived from non-human cell lines (e.g., murine or Chinese hamster ovary (CHO) cells). Even non-human-like glycan epitopes (e.g., β1,2-xylose in plant-derived biologics) may cause hypersensitivity. Regulatory agencies require a thorough glycan immunogenicity risk assessment to ensure patient safety. Advanced glycoengineering and cell line optimization (e.g., using glycoKO or glycohumanized systems), for instance, might help minimize these risks.
  • Biological Activity – Glycosylation directly impacts protein-receptor interactions, altering binding affinity and downstream signaling. For example, IgG Fc-glycosylation determines whether an antibody activates or suppresses inflammation—afucosylated IgGs show stronger binding to FcγRIIIa, enhancing NK cell-mediated cytotoxicity, while sialylated IgGs promote anti-inflammatory effects via FcγRIIb. Similarly, EGFR and HER2 (key oncology targets) exhibit altered signaling when glycosylation sites are mutated. Even G-protein-coupled receptors (GPCRs) rely on glycans for proper folding and ligand recognition. By performing a thorough glycan analysis, researchers can correlate specific glycoforms with biological activity, enabling structure-function optimization for next-gen biologics.

Glycan profiling is a cornerstone in the development and quality control of biopharmaceuticals – regulatory agencies (FDA, EMA) require comprehensive glycan characterization for biopharmaceutical approval processes- with applications spanning multiple stages of drug development.

In this context, there are certain Key Applications of Glycan Profiling that cannot be missed:

  1. Monoclonal Antibodies (mAbs): One of the most critical areas is in monoclonal antibody (mAb) therapeutics, where glycosylation—particularly at the Fc region—directly influences effector functions such as antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). For example, the presence of core fucosylation reduces ADCC, while bisecting GlcNAc and high-mannose glycans can enhance it. This has led to targeted glycoengineering approaches to optimize therapeutic efficacy, as seen in next-generation antibodies like obinutuzumab, which exhibits reduced fucosylation for improved cancer cell targeting.
  2. Recombinant Proteins: Beyond mAbs, recombinant proteins such as erythropoietin (EPO) and clotting factors (e.g., Factor VIII) rely heavily on proper glycosylation for stability and biological activity. In EPO, as previously mentioned, sialic acid-capped N-glycans are essential for extending serum half-life by preventing rapid clearance via the asialoglycoprotein receptor.
  3. Biosimilars: Similarly, biosimilar development demands rigorous glycan comparison to ensure structural and functional equivalence to reference products—a requirement enforced by regulatory agencies like the FDA and EMA.
  4. Vaccines Additionally, vaccine development benefits from glycan analysis and profiling, as viral surface glycoproteins (e.g., HIV’s gp120 or SARS-CoV-2’s Spike protein) often use glycans to evade immune detection. Mapping these glycosylation patterns helps in designing more effective vaccines by either mimicking natural glycosylation or modifying it to enhance immunogenicity.
Glycan Analysis and Profiling by MALDI-TOF
Glycan Profiling Spectra: Glycan Analysis by MALDI-TOF

Asparia Glycomics: Your Partner in Glycan Analysis

At Asparia Glycomics, we offer state-of-the-art glycan analysis services using cutting-edge technologies combined and/or separately, such as:

  • Liquid Chromatography (HPLC/UPLC)
  • Nuclear magnetic resonance (NMR)
  • Mass Spectrometry (MS), MALDI-TOF

Our services help biopharma companies:


✔ Ensure batch-to-batch consistency
✔ Optimize glycoengineering strategies
✔ Meet regulatory requirements
✔ Accelerate drug development timelines

Why Choose Asparia Glycomics?

  • We are experts in Glycoscience – Asparia Glycomics has a formidable team of chemists and molecular biologists who can understand the client needs, priorities and select and/or combine both types of glycan analysis in record time.
  • High-Throughput Capabilities – Fast, accurate results for large-scale studies.
  • Custom Solutions – Tailored approaches for unique project needs.

Our projects are outstanding for fixed rates, flexible communication including consultation, and fast turnaround time.

Conclusion

Glycan analysis is indispensable for developing safe, effective biopharmaceuticals. Asparia Glycomics provides cutting-edge glycan analysis services to support drug development, ensuring compliance and optimal product performance.

If you´d like to have a meeting with us 📩 Contact us today to learn how our glycan profiling solutions can enhance your biopharmaceutical projects!

Bibliography – Glycan Analysis:

Anthony, R. M. et al. (2008)
Recapitulation of IVIG anti-inflammatory activity with a recombinant IgG Fc
Science. DOI: 10.1126/science.1154315

Ashwell, G. & Harford, J. (1982)
Carbohydrate-specific receptors of the liver
Annual Review of Biochemistry. DOI: 10.1146/annurev.bi.51.070182.002531

EMA (2017)
Immunogenicity assessment of therapeutic proteins
EMA/CHMP/BMWP/14327/2006. Link

FDA (2020)
Immunogenicity Testing of Therapeutic Protein Products
Link

Ghaderi, D. et al. (2010)
Implications of the presence of N-glycolylneuraminic acid in recombinant therapeutic glycoproteins
Nature Biotechnology. DOI: 10.1038/nbt.1651

Hamilton, S. R. et al. (2006)
Humanization of yeast to produce complex terminally sialylated glycoproteins
Science. DOI: 10.1126/science.1130256

Higel, F. et al. (2016)
N-glycosylation heterogeneity and the influence on structure, function and pharmacokinetics of monoclonal antibodies and Fc fusion proteins
European Journal of Pharmaceutics and Biopharmaceutics. DOI: 10.1016/j.ejpb.2016.05.022

ICH Q6B (1999)
Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological Products
Link

Jefferis, R. (2017)
Glycosylation as a strategy to improve antibody-based therapeutics
Nature Reviews Drug Discovery. DOI: 10.1038/nrd.2016.246

Liu, Y. C. et al. (2011)
The role of N-glycosylation in EGFR signaling and trafficking
Glycobiology. DOI: 10.1093/glycob/cwr080

Macher, B. A. & Galili, U. (2008)
The Galα1,3Galβ1,4GlcNAc-R (α-Gal) epitope: a carbohydrate of unique evolution and clinical relevance
Biochimica et Biophysica Acta (BBA)-General Subjects. DOI: 10.1016/j.bbagen.2007.12.001

Moremen, K. W. et al. (2012)
Vertebrate protein glycosylation: diversity, synthesis and function
Nature Reviews Molecular Cell Biology. DOI: 10.1038/nrm3383

Reusch, D. & Tejada, M. L. (2015)
Fc glycans of therapeutic antibodies as critical quality attributes
Glycobiology. DOI: 10.1093/glycob/cwv037

Reusch, D. et al. (2015)
Glycosylation of biosimilars: Current challenges and analytical solutions
mAbs. DOI: 10.1080/19420862.2015.1028634

Roopenian, D. C. & Akilesh, S. (2007)
FcRn: the neonatal Fc receptor comes of age
Nature Reviews Immunology. DOI: 10.1038/nri2157

Sakaue, H. et al. (2020)
Glycosylation of G protein-coupled receptors: roles in receptor pharmacology and disease pathogenesis
British Journal of Pharmacology. DOI: 10.1111/bph.14984

Shields, R. L. et al. (2002)
Lack of fucose on human IgG1 N-linked oligosaccharide improves binding to human FcγRIII and antibody-dependent cellular toxicity
Journal of Biological Chemistry. DOI: 10.1074/jbc.M202069200

Yang, Z. et al. (2015)
Engineered CHO cells for production of diverse, homogeneous glycoproteins
Nature Biotechnology. DOI: 10.1038/nbt.3280

What are the Mass Spectrometry-based techniques for Glycan Analysis?

Posted on by Asparia Glycomics

Glycan analysis using mass spectrometry (MS) is a crucial technique for elucidating, characterizing, and quantifying glycans in biological samples. Mass spectrometry-based glycomics allows for high-throughput profiling of N-glycans, O-glycans, glycopeptides, and glycolipids with exceptional sensitivity and accuracy. Common MS techniques employed in glycan analysis include matrix-assisted laser desorption/ionization (MALDI-MS), electrospray ionization (ESI-MS), ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS), and tandem mass spectrometry (MS/MS). Each of these methods has specific advantages and limitations.

Pros and Cons of each different MS-based techniques

MALDI-MS

MALDI-MS enables rapid, high-throughput, and cost-effective screening of large numbers of samples when associated with automated sample preparation. However, its lower sensitivity in detecting minor glycan species and the impossibility of distinguishing glycan isomers can limit its applicability for complex samples.

ESI-MS

ESI-MS, especially when paired with UHPLC, offers enhanced sensitivity and allows for detailed structural characterization of glycan compositions, linkage patterns, and structural isomers. When combined with hydrophilic interaction liquid chromatography (HILIC), glycans (including isomeric forms) are separated based on their interactions with the column, which improves the accuracy of identification. This method is particularly effective for analyzing complex samples such as tissue, biofluids like plasma, or cell extracts.

TOF-MS

Glycan analysis using mass spectrometry involves derivatization techniques, such as permethylation or fluorescent labeling (e.g., procainamide), to improve glycan detection and ionization efficiency. Time-of-flight mass spectrometry (TOF-MS), which is a type of high-resolution mass spectrometry (HRMS), provides accurate mass determinations within a 15 ppm error range, making it highly effective for profiling glycan structures. For example, the signals of closely related glycans can be completely separated, even in cases of chromatography co-elution, something that non-mass-based detectors, such as UV or fluorescence detectors, cannot achieve.

Tandem MS

Tandem MS (MS/MS) techniques, such as higher-energy collisional dissociation (HCD), provide in-depth fragmentation data for glycan structural analysis. These methods are crucial for distinguishing isomeric glycans and elucidating complex glycosylation patterns. However, the fragmentation patterns of glycans can be challenging to interpret due to the complexity of glycosidic and cross-ring cleavages, necessitating specialized software tools for data analysis.

MS-based applications

Mass spectrometry-based glycomics has numerous applications, particularly in biopharmaceutical development. Glycan profiling is essential for the quality control of monoclonal antibodies (mAbs), recombinant glycoproteins, and vaccines. In glycoengineering, MS-based analysis aids in optimizing glycosylation to improve drug stability, efficacy, and immunogenicity. Additionally, glycan mass spectrometry plays a significant role in biomarker discovery, helping to identify disease-associated glycosylation patterns linked to cancer, autoimmune disorders, and infectious diseases. Quantitative glycomics approaches that use stable isotope-labeled glycan standards enable the accurate quantification of glycans in complex biological matrices and facilitate comparative studies between diseased and healthy states.

As glycobiology becomes increasingly important in biomedical research, ongoing improvements in analytical workflows, data interpretation, and automation enhance the efficiency and reproducibility of glycan characterization. The combination of mass spectrometry with complementary techniques such as chromatography and bioinformatics is driving innovation in glycomics, ultimately supporting the development of novel therapeutics, diagnostics, and personalized medicine strategies.

Discover what Asparia does for your glycan analysis needs

In Asparia Glycomics, we are experts in glycan analysis, and we have access to last-generation equipment and instruments to apply MALDI, ESI, Tandem, and TOF-MS to a broad variety of samples. This fact, combined with the expertise in the scientific teams, transforms Asparia into a reliable partner to give answers to your research and R&D questions related to the glycans present in any of your samples. We also offer a door-to-door sample delivery service to ease and smooth the whole analytical process. We take pride in providing a very thorough report after analysis, compliant with the highest publication standards, and if you have any doubts, we are always available to further explain and discuss your valuable results.  

CarboQuant: The Cutting-Edge Glycan Analysis Technology Transforming Disease Research

Posted on by Asparia Glycomics

Glycans play a crucial role in many biological processes, from cell signaling and immune response to disease progression. However, accurately quantifying these complex carbohydrates has long been a challenge in biomedical research. That’s where CarboQuant comes in—our proprietary technology designed to provide absolute glycan quantification with unparalleled precision. By leveraging synthetic ¹³C-labeled N-glycans and mass spectrometry, CarboQuant enables researchers to uncover glycan biomarkers linked to diseases such as cancer, Alzheimer’s, and diabetes, driving forward innovation in diagnostics and therapeutics.

The Significance of Glycans in Disease Research

Glycans, complex carbohydrates present on the surfaces of cells and proteins, play pivotal roles in numerous biological processes, including cell signaling, immune response modulation, and protein stability. Alterations in glycan structures are often associated with various diseases, such as cancer, Alzheimer’s, and diabetes. Therefore, precise glycan analysis is essential for understanding disease mechanisms and developing effective diagnostics and therapeutics.

CarboQuant Technology: Revolutionizing Glycan Quantification

CarboQuant stands out as a patented technology developed by Asparia Glycomics, utilizing synthetic 13C-labeled N-glycans for absolute quantification by mass spectrometry. This approach ensures high precision in glycan measurement, which is crucial for identifying disease-specific glycan biomarkers. The technology’s robustness and accuracy have made it a valuable asset in biopharmaceutical research, particularly in quality control and biomarker discovery.

Applications of CarboQuant in Disease Research

Cancer Diagnostics and Prognostics

Aberrant glycosylation patterns are a hallmark of many cancers. CarboQuant enables the detailed profiling of these glycan alterations, facilitating the identification of potential biomarkers for early cancer detection and prognosis. By providing absolute quantification, researchers can distinguish subtle differences in glycan expression that may correlate with tumor progression or response to therapy.

Alzheimer’s Disease Research

In neurodegenerative diseases like Alzheimer’s, glycosylation changes can affect protein aggregation and neuronal function. Utilizing CarboQuant, scientists can accurately measure these glycan modifications, shedding light on disease pathways and identifying novel therapeutic targets. This precise quantification is essential for developing strategies to mitigate glycan-related pathogenic mechanisms in Alzheimer’s disease.

Diabetes and Metabolic Disorders

Glycans play a significant role in insulin activity and glucose metabolism. Alterations in glycan structures can influence insulin resistance and the development of diabetes. CarboQuant might facilitate the monitoring of these glycan changes, aiding in the understanding of disease progression and the development of glycan-targeted therapies.

Integration with Industry-Standard Platforms

Asparia Glycomics has ensured that CarboQuant technology is compatible with widely used analytical platforms for Mass Spectrometry and MALDI. This compatibility allows for seamless integration into existing workflows, enabling researchers to adopt CarboQuant without the need for extensive modifications to their current systems. The result is a streamlined process for glycan analysis, enhancing efficiency and accuracy in research applications.

Custom Glycan Synthesis Services

Recognizing that research needs can vary, and some N-glycan standards are not always commercially available, Asparia Glycomics offers custom glycan synthesis services, including SIL (Stable Isotope Labeled)-standards. This flexibility allows scientists to obtain specific glycan structures tailored to their research objectives, whether for developing new therapeutics or exploring unique disease mechanisms. The ability to customize glycan standards ensures that researchers have the precise tools necessary for their specific applications.

CarboQuant
Figure 1. Selection of target compounds. Source: Own elaboration.

Global Impact and Collaborations

Operating on an international scale, Asparia Glycomics collaborates with pharmaceutical companies, biotech firms, and academic institutions. Their technologies and services have been instrumental in advancing glycomics research worldwide, contributing to significant progress in understanding and treating complex diseases. These collaborations have facilitated the translation of glycan research into clinical applications, ultimately benefiting patient care and therapeutic development.

Future Directions

Asparia Glycomics is committed to expanding its technological capabilities and service offerings. Plans include a wider implementation of the CarboQuant technology and the use of the standards for a better exploration of glycan-related biomarkers and in a broader range of diseases. By staying at the cutting edge of glycoscience, Asparia Glycomics aims to provide researchers and scientists with the necessary tools to drive innovations in diagnostics and therapeutics.

Conclusion

The CarboQuant technology represents a significant advancement in the field of glycan analysis. Its precise quantification capabilities are invaluable for disease research, offering insights into glycan-related mechanisms across various pathologies. As the scientific community continues to uncover the complexities of glycosylation in health and disease, tools like CarboQuant will be essential in translating this knowledge into tangible medical advancements.

References

Aizpurua-Olaizola, O., Sastre Toraño, J., Falcon-Perez, J. M., Williams, C., Reichardt, N., & Boons, G.-J. . (2018). Mass spectrometry for glycan biomarker discovery. TrAC Trends in Analytical Chemistry, 100, 7–14. https://doi.org/10.1016/j.trac.2017.12.015

Etxebarria, J., & Reichardt, N.-C. (2016). Methods for the absolute quantification of N-glycan biomarkers. Biochimica et Biophysica Acta (BBA) – General Subjects, 1860(8), 1676–1687. https://doi.org/10.1016/j.bbagen.2016.03.003

Echeverria, B., Etxebarria, J., Ruiz, N., Hernandez, Á., Calvo, J., Haberger, M., Reusch, D., & Reichardt, N.-C. (2015). Chemo-Enzymatic Synthesis of 13C Labeled Complex N-Glycans As Internal Standards for the Absolute Glycan Quantification by Mass Spectrometry. Analytical Chemistry, 87(22), 11460–11467. https://doi.org/10.1021/acs.analchem.5b03135

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In conversation with Leyre Aramendia, Senior Research Biochemist

Posted on by Asparia Glycomics

Leyre joined Asparia in 2019 as a Research Biochemist, working in both Glycobiology and Analytical Chemistry. Before that, she gained experience in biopharmaceutical production at a CRO. She specializes in the enzymatic synthesis of complex oligosaccharides and glycopeptides, as well as the production and analysis of synthetic glycans. She’s also involved in N- and O-glycan analysis and the isolation of glycans from natural sources. Leyre holds a Biochemistry degree from the Universidad Autónoma de Madrid and a Master’s in Bioinformatics and Biostatistics from the Universitat Oberta de Catalunya.

In this interview, we’ll be diving into the world of enzymatic synthesis and glycan analysis, breaking down why they’re so essential in biotechnology. We’ll talk about how enzymatic approaches offer significant advantages over traditional chemical methods, making glycan synthesis more efficient and precise.

Beyond that, we’ll explore some of the biggest challenges in the field, as well as the latest advancements that are helping to overcome them. And, of course, we’ll discuss the crucial role glycans play in biotechnology—especially in terms of biomolecule stability and their growing importance in therapeutic applications.

Introduction to Enzymatic Synthesis and Glycan Analysis

Your research focuses on enzymatic synthesis and glycan analysis. For those who may not be familiar with these concepts, could you explain what they entail and why they are so important in the field of biotechnology?

Of course! Enzymatic synthesis uses specialized enzymes to build or modify glycans—complex carbohydrate molecules—with high precision and efficiency. Enzymes like glycosyltransferases and glycosidases help assemble glycan structures with incredible specificity. This is crucial for developing glyco-conjugates used in vaccines, cancer therapies, biotechnology, and disease diagnostics.

Glycan analysis, on the other hand, studies the structure, composition, and function of glycans attached to proteins and lipids. Since glycans play key roles in biological processes, understanding them helps advance diagnostics, therapies, and vaccine development. Together, these fields drive innovation in personalized medicine and sustainable industrial processes.

Enzymatic synthesis has gained significant importance in recent years due to its specificity and efficiency. What are the main advantages of using enzymes for glycan synthesis compared to traditional chemical methods?

Unlike chemical synthesis, which often requires harsh conditions and can produce unwanted byproducts, enzymatic synthesis is highly specific and efficient. Enzymes act as biological catalysts, allowing reactions to occur under mild conditions while minimizing side reactions. This results in purer, more consistent glycans, making enzymatic synthesis a superior alternative for many applications.

Regarding glycan analysis, Asparia Glycomics utilizes advanced technologies for their characterization. What methodologies are employed in the laboratory to conduct this analysis, and what kind of information can be obtained from the samples?

Developing new methodologies for glycan synthesis and analysis is challenging due to the structural complexity and heterogeneity of glycans. Their highly branched and diverse nature makes both synthesis and characterization difficult, requiring precise enzymatic or chemical strategies. However, enzyme availability and specificity often limit synthesis efficiency, while scalable production remains a challenge.

On the analytical side, the lack of standardized databases and the complexity of biological samples make glycan identification and quantification difficult. Advanced techniques like mass spectrometry (MS) and liquid chromatography (LC) are essential but require specialized expertise and equipment, increasing costs and limiting accessibility. Additionally, integrating glycan analysis with other omics fields, such as genomics and proteomics, remains a major hurdle due to the absence of standardized cross-disciplinary tools.

Challenges and Methodological Development

Your research sits at the intersection of chemistry, biology, and biotechnology. What are the main challenges you encounter in developing new methodologies for glycan synthesis and analysis?

The biggest challenge in my opinion comes from the structural complexity of glycans—as previously said, they’re highly branched and diverse, making both synthesis and analysis difficult. On the synthetic side, enzymatic synthesis is limited by enzyme availability and specificity, making the scaling up process challenging.

Regarding the techniques, advanced techniques like MS and LC require specialized expertise and expensive equipment, which limits accessibility. Also, integrating glycan analysis with other omics fields, like genomics and proteomics, is another challenge due to the lack of standardized tools.

What are the most commonly used enzymes in enzymatic glycan synthesis, and what are their primary functions?

The most used enzymes in enzymatic glycan synthesis are glycosyltransferases (GTs). These enzymes are central as they transfer sugar units to acceptor molecules, building the glycan chains by forming specific glycosidic linkages. While GTs are the primary tools for constructing glycans, other types of enzymes also play essential roles in modifying and fine-tuning glycan structures.

For example, glycosidases are used to break or modify glycosidic bonds, removing or altering sugar residues. Epimerases change the stereochemistry of sugars, sulfotransferases add sulfate groups to glycans, and sialyltransferases transfer sialic acid to glycan chains. Though glycosyltransferases are the most widely employed, these other enzymes are equally important for adjusting the structure and function of glycans in a precise and controlled manner.

What are the current limitations of enzymatic glycan synthesis, and what strategies are being implemented to address them?

I´d say that enzymatic glycan synthesis faces several challenges, including limited enzyme specificity, scalability issues, glycan heterogeneity, and substrate limitations.

Strategies to overcome these challenges may include enzyme engineering to improve specificity, the development of multi-enzyme systems for more efficient synthesis, and advanced analytical techniques to monitor and control glycan structures. Additionally, efforts are focused on improving the availability and cost-effectiveness of sugar donors, standardizing synthesis protocols, and utilizing more biologically relevant environments, such as cell-free systems and microbial platforms, for production.

Structural and Functional Aspects of Glycans

How does the three-dimensional structure of a glycan influence its interaction with enzymes during synthesis?

It is a very interesting question: the 3D structure of a glycan determines how well it fits into an enzyme’s active site. Steric hindrance from bulky or branched glycans can slow down or block enzyme binding. Conversely, certain structural features can enhance enzyme recognition, ensuring efficient catalysis.

Is there a connection between enzymatic synthesis and glycoprotein bioengineering for therapeutic applications?

Enzymatic synthesis is essential for glycoprotein bioengineering because it allows precise control over glycosylation patterns, which directly impacts a protein’s stability, function, and immune response. This is crucial in developing biopharmaceuticals, vaccines, and targeted therapies. By fine-tuning glycan structures, we can optimize therapeutic proteins for better efficacy and aiming too for fewer side effects.

How does glycosylation impact the stability and function of biomolecules, and how can it be optimized through enzymatic synthesis?

As I just mentioned, glycosylation is critical for the stability, function, and activity of biomolecules, particularly proteins. The addition of specific sugar chains to proteins can enhance their stability by protecting them from degradation. It can also improve their solubility and influence their folding and conformation. But interestingly, glycosylation also affects protein interactions, recognition, and immune responses, which are crucial for therapeutic efficacy. Through enzymatic synthesis, glycosylation might be precisely controlled to optimize these properties or modifying sugar residues in a targeted manner, to ensure that the biomolecule of interest exhibits specific characteristics for its intended application, such as in biopharmaceuticals or vaccines.


Reflecting on 2024: A year of growth, innovation, and collaboration

Posted on by Asparia Glycomics

The end of the year is almost here, and at Asparia Glycomics, we want to take a moment to reflect on the remarkable journey we’ve shared with our clients and partners throughout 2024. It’s been a year filled with innovation, growth, and meaningful connections that have inspired us to push boundaries and reach new milestones. Your trust, collaboration, and continued support have been the driving forces behind our achievements this year, and for that, we are truly grateful. Together, we’ve created impactful solutions and laid the groundwork for an even brighter future ahead.

A year of innovation, ambitious projects and results

This year has been a testament to innovation, growth, and shared success. At Asparia, we took on 25 unique projects, with particular emphasis on glycan analysis—a field where each sample is a world of complexity and high value for R&D. Our team has relished the challenge of delivering key compounds and results that empower our clients’ goals.

We’re proud to have collaborated with 30 different clients, spanning both the private and academic sectors. These partnerships have allowed us to push boundaries, refine our expertise, and make a meaningful impact in the Pharma and Biotech industries.

Strengthening connections across Europe

In 2024, we strengthened our presence at key European events, attending the Swiss Biotech Day and BioEurope, while also sponsoring and participating in the VII Iberian Carbohydrate Meeting and Glycobasque7. These platforms offered invaluable opportunities to understand the evolving needs of our industry, reinforce relationships, and cement our role as a trusted reference in glycomics for both industry and academia.

innovation
Image 1. An overview of the BioEurope event.

We’re also thrilled to have joined AseBio as a member this year, underscoring our commitment to playing a larger role in Spain’s biotechnology landscape. This step not only enhances our visibility but also positions us to actively contribute to the sector’s growth and innovation.

Growth from within: Welcoming new team members

Growth has come from within, too. In 2024, we welcomed two key additions to our team: Jaime Escriba, as Business Development Specialist, and Oscar Lobera, who stepped in as our CEO, bringing fresh perspectives and leadership to help us achieve our ambitious goals.

Looking ahead: Ambitions for 2025

Looking ahead to 2025, our vision is clear: we aim to consolidate Asparia Glycomics as a global leader in carbohydrate synthesis and as a trusted partner in glycoanalysis. To support this growth, we are embarking on an exciting expansion project, scaling up our facilities at the Gipuzkoa Technology Park. This investment will further enhance our technical capabilities and allow us to better serve our clients across the Pharma and Biotech markets.

A heartfelt thank you

As we close this chapter and prepare for the opportunities ahead, we reflect on the incredible moments that defined this year. Every challenge overcome, every milestone reached, and every collaboration forged has been a testament to the collective effort of our team and the unwavering support of our partners.

Looking ahead to 2025, we are fueled by a renewed sense of purpose and the drive to push boundaries in the field of glycomics. We envision a year filled with transformative ideas, deeper connections, and groundbreaking advancements. None of this would be possible without you—our clients, partners, and friends—who inspire us to aim higher every day.

From exciting research breakthroughs to innovative collaborations, we are determined to make the upcoming year even more impactful. Your support motivates us to constantly evolve and excel.

From all of us at Asparia Glycomics, thank you for your trust and partnership. Let’s continue this journey together and make 2025 a year to remember.

Wishing you a joyful holiday season and a prosperous New Year!

Microbiota Glycans: Shaping Health, Microbes and Response of the Immune System

Posted on by Asparia Glycomics

The influence of glycans on the human gut microbiota is a central topic in contemporary glycoscience. These type of carbohydrates in this environment are not only essential for microbial colonization but also serve important regulatory functions that contribute to immune protection. In this article, we will explore the fundamental roles of glycans, present intriguing case studies, and highlight two practical applications.

Our discussion is informed by the outstanding work of the research team behind a comprehensive study published in May of this year. This study involves scientists from the University of Birmingham and the University of Porto and offers a broad perspective on the current state of the field, addressing key aspects of glycobiology and referencing a wide array of relevant research that has shaped our understanding of the microbiota-glycan connection.

The Glycan Landscape within the Human Microbiota

The collection of lipids and proteins decorated with diverse glycans on the surface of the gut mucosa is called glycocalyx and serves as a major interface between the intestinal lining and microorganisms, as well as with the host’s immune system.  Below, we highlight the key structures within the glycocalyx to illustrate how these carbphydrates shape the gut environment, drive microbial interactions, and influence immune system regulation.

glycans

Illustration of a healthy colonic human glycocalyx, from The role of glycans in health and disease: Regulators of the interaction between gut microbiota and host immune system. In Seminars in Immunology (Vol. 73, p. 101891). Academic Press.

  • Glycoproteins and Glycolipids: The epithelial cell membrane expresses highly diverse glycoproteins and glycolipids, which are decorated with N-linked and O-linked glycans. These carbohydrates, when recognized by immune cells, can dictate the nature of the immune response—either pro-inflammatory or anti-inflammatory.
  • Glycan-Binding Proteins (GBPs): Immune cells produce GBPs, such as galectins, siglecs, and C-type lectins, which «read» the glycan code expressed by both host cells and microbes.
  • Glycosaminoglycans (GAGs): Chondroitin sulfate, heparan sulfate, and hyaluronan are also important glycans of the glycocalyx. They serve as nutrient sources for gut microbes, such as Bacteroides thetaiotaomicron, highlighting their role in microbial metabolism and colonic health.
  • Mucins (MUC1–22): Mucins form the protective mucus layer overlaying the glycocalyx. As an example, it has been shown that MUC1 functions as a decoy to distract and confuse Helicobacter pylori during infection.
  • Carbohydrate-Active Enzymes (CAZymes): Gut microbes produce CAZymes like glycoside hydrolases (enzymes depicted as «scissors») to degrade complex glycans into short-chain fatty acids (SCFAs). SCFAs act as essential nutrients for colonocytes and help maintain gut homeostasis.

The Influence of Glycans: Intriguing Case Studies

Several studies have identified the significant role of certain glycans within our microbiota in regulating immune system responses and determining which microbes colonize our gut, ultimately influencing our health and susceptibility to various diseases.

A noteworthy example of this interaction can be seen in research on infants who are either breastfed or formula-fed. Studies reveal that human milk oligosaccharides (HMOs) play a crucial role in fostering the growth of Bifidobacterium longum subsp. infantis, a beneficial bacterium that thrives predominantly in the gut of breastfed infants.

In a more surprising twist, some pathogens have evolved a strategy known as glycan mimicry, where they display glycans on their surface that immune cells fail to recognize as harmful, allowing them to evade detection. A striking example is Neisseria gonorrhoeae, the bacteria responsible for gonorrhea, which incorporates sialic acid from the host onto its lipooligosaccharides, effectively camouflaging itself from the immune system.

Targeting the Glyco-Microenvironment: Two Practical Applications

Several approaches are being explored to target the intestinal glyco-microenvironment, including the enhancement of the mucosal glycome through glycan supplementation or by modifying the activity of CAZymes. Two recent studies have made significant strides in translating these concepts into practical applications.

One study, published in Nature, involves researchers from Washington University School of Medicine and the International Centre for Diarrhoeal Disease Research in Dhaka, Bangladesh. The team identified key bioactive glycan structures, contributing to the development of their MDCF-2 project. This therapeutic food is designed to help restore the gut microbiota of malnourished children, offering hope for improving their gastrointestinal health.

Another study, published in the Proceedings of the National Academy of Sciences, focuses on how gut microbes process plant-derived N-glycans. By identifying specific enzymes that can modify and analyze these N-glycan structures, the research aims to apply these findings to medical and industrial fields. One of the possible applications is to reduce the risk of allergic reactions to foods and medications by leveraging this knowledge.

Conclusions

The intricate interactions between glycans and gut microbes are crucial for understanding and maintaining gut health, as well as for treating immune diseases and gastrointestinal disorders. At Asparia Glycomics, we specialize in supporting cutting-edge research in this field. Whether you’re developing HMOs for synthetic milk or unraveling the composition and structure of bioactive oligosaccharides and polysaccharides, our expertise in glycan synthesis and analysis is here to guide you. We have a proven track record of assisting projects in gut microbiota research and development.

As advances in glycomics continue to unfold, they are unlocking transformative medical applications, from restoring microbiomes in malnourished children to mitigating allergic reactions through glycan engineering. These special carbohydrates undoubtedly hold the key to groundbreaking solutions in human health, and we at Asparia Glycomics are ready to partner with you on this exciting journey. Reach out to explore how we can support your innovation.

References

  • Crouch, L. I., Rodrigues, C. S., Bakshani, C. R., Tavares-Gomes, L., Gaifem, J., & Pinho, S. S. (2024, May). The role of glycans in health and disease: Regulators of the interaction between gut microbiota and host immune system. In Seminars in immunology(Vol. 73, p. 101891). Academic Press.
  • Crouch, L. I., Urbanowicz, P. A., Baslé, A., Cai, Z. P., Liu, L., Voglmeir, J., … & Bolam, D. N. (2022). Plant N-glycan breakdown by human gut Bacteroides. Proceedings of the National Academy of Sciences, 119(39), e2208168119.
  • Hibberd, M. C., Webber, D. M., Rodionov, D. A., Henrissat, S., Chen, R. Y., Zhou, C., … & Gordon, J. I. (2024). Bioactive glycans in a microbiome-directed food for children with malnutrition. Nature, 625(7993), 157-165.
  • Wang, Y., Ze, X., Rui, B., Li, X., Zeng, N., Yuan, J., … & Li, M. (2021). Studies and application of sialylated milk components on regulating neonatal gut microbiota and health. Frontiers in Nutrition, 8, 766606.

The Path of Glycan-Based Biomarkers to Transform Cancer Diagnostics and Therapeutics

Posted on by Asparia Glycomics

The quest to improve cancer diagnostics and therapeutics remains at the forefront of modern science, and glycoscience may hold the potential for groundbreaking progress. Why focus on glycoscience? Because changes in glycosylation—a fundamental post-translational modification—are closely associated with cancer progression. These glycosylation alterations often lead to unique glycan structures that can serve as reliable biomarkers for various cancers. Indeed, glycan-based biomarkers have been identified in several cancer types, including breast, ovarian, colorectal, prostate, pancreatic cancers, as well as lymphoma and myeloma, promising to enhance early detection and personalized treatment approaches.

But how close are we to fully implementing these findings to transform patient care? What advancements have already been made, and which techniques show the most promise? In this article, we delve into the current landscape of glycan-focused cancer diagnostics and therapeutics, spotlight two recent studies with compelling results, and share insights from Antoine Lesur, analytical scientist and Head of Analytics at Asparia Glycomics.

The Need for Standardization and Multidisciplinary Efforts

The well-established link between aberrant glycosylation and the initiation and progression of numerous cancers has propelled cancer glycomics research towards identifying specific glycan biomarkers. These biomarkers hold the potential to revolutionize cancer diagnosis and personalized treatment. Over the past few decades, efforts have largely focused on advancing technologies that allow the observation of large-scale glycosylation changes. Additionally, considerable progress has been made in connecting glycans to their protein carriers and exploring how these modifications impact intracellular signaling and cellular functions.

Despite these advancements, we are still at the early stages of fully deciphering glycosylation’s complex role in cancer. One of the critical challenges hindering successful biomarker discovery is the lack of standardized and accessible protocols for glycan sample preparation, data acquisition, and analysis. Therefore, collaborative efforts with experts across pertinent fields are essential. As Antoine Lesur emphasizes, «During the steps of biomarker discovery, verification, and validation, the choice of samples must be made in advance with the help of clinicians and statisticians to ensure that the data will be relevant.»

In addition, inter-subject variability studies should be integrated into glycan biomarker research. As outlined in our publication «Mass Spectrometry for Glycan Biomarker Discovery,» to ensure accuracy, any glycan biomarker study must include an analysis on variability between the genetic and demographic differences among individuals and due to specific pathological conditions.

Mass Spectrometry Techniques Driving the Present and Future

Various glycan analysis techniques can be applied to biomarker discovery, with mass spectrometry (MS) being an indispensable tool due to its precision in mass measurement for compositional analysis and its ability for structural identification.

For N- glycan analysis, a commonly employed MS technique is MALDI (matrix-assisted laser desorption/ionization), where a sample on a solid support can be combined with a matrix and undergo ionization. This technique is applied to the analysis of tissues of clinical samples in cancer.

For classic glycomics, it is also common to integrate complementary techniques. Mass spectrometry methods allow the possibility to couple with separation techniques like liquid chromatography, capillary electrophoresis and ion mobility. These combined techniques offer significant potential for separating isomers, which could be crucial for discovering new biomarkers. They are also regularly employed for quality control in biotherapeutics used in the treatment of cancer.

Breakthrough Studies of 2024 in Cancer Glycomics Research

In May of this year, researchers from Ivanov Lab at Northeastern University published a study in Nature Communications reporting unique results. They successfully applied capillary electrophoresis followed by mass spectrometry to analyze a single still-living human cell. Before this, nobody had examined, measured, and determined the structure of whole proteins and native glycans from an individual cell. This marked a relevant advanced towards diagnostics tests for a variety of diseases, including cancer.

Just a month later, a team from the Australian Institute for Bioengineering and Nanotechnology introduced another innovative technology in Advanced Science, that lead to significant results. They developed a nanodevice capable of analyzing glycopatterns of small extracellular vesicles (sEVs) released by lung cancer cells. In a clinical study of 40 patients, this nanotechnology distinguished early-stage malignant lung nodules from benign ones, demonstrating the potential of profiling small EV glycans for noninvasive lung cancer diagnostics and prognostics.

Tumor-Associated Carbohydrate Antigens: Potential Cancer Biomarkers

Among the diverse range of cancer biomarkers, Tumor-Associated Carbohydrate Antigens (TACAs) have drawn attention from researchers: «TACAs are glycan structures that are abnormally expressed in cancer cells. They play significant roles in tumor growth, metastasis, and immune evasion, which makes them potential biomarkers for cancer diagnosis and potential therapeutic targets», explains Antoine.

Some common TACAs are Thomsen-nouvelle antigen (Tn), sialyl Thomsen-nouvelle antigen (STn) and sialyl Lewis antigens. Tn and STn antigens have been identified in nearly all types of human carcinomas and are highly overexpressed in breast, colon, lung, pancreatic, and prostate cancers.

TACAs are commonly identified using chromatographic techniques combined with mass spectrometry. This process starts with extracting glycoproteins from serum or plasma, followed by purification involving antibodies or lectins, and then the glycans are detached from the peptide backbone through chemical or enzymatic treatments. Afterwards, the glycans are separated based on their structure polarity and size, allowing direct analysis via native MS.

Conclusions

Significant progress has been made with analytical techniques like mass spectrometry and the study of tumor-associated carbohydrate antigens (TACAs). However, challenges remain—particularly around standardization and the need for a stronger multidisciplinary approach.

To truly position glycoscience as a cornerstone of personalized medicine, greater investment, specialized expertise, and precision are essential. Asparia’s Glycan Analysis Service, led by Antoine Lesur, supports this vision by offering precise mapping of glycosylation patterns, enabling the accurate identification and synthesis of TACAs to propel your research and development forward.

References

  • Aizpurua-Olaizola, O., Toraño, J. S., Falcon-Perez, J. M., Williams, C., Reichardt, N., & Boons, G. J. (2018). Mass spectrometry for glycan biomarker discovery. TrAC Trends in Analytical Chemistry, 100, 7-14.
  • Doud, E. H., & Yeh, E. S. (2023). Mass spectrometry-based glycoproteomic workflows for cancer biomarker discovery. Technology in Cancer Research & Treatment, 22, 15330338221148811.
  • Marie, A. L., Gao, Y., & Ivanov, A. R. (2024). Native N-glycome profiling of single cells and ng-level blood isolates using label-free capillary electrophoresis-mass spectrometry. Nature Communications, 15(1), 3847.
  • Puiu, M., Nativi, C., & Bala, C. (2023). Early detection of tumour-associated antigens: Assessment of point-of-care electrochemical immunoassays. TrAC Trends in Analytical Chemistry, 160, 116981.
  • Zhou, Q., Niu, X., Zhang, Z., O’Byrne, K., Kulasinghe, A., Fielding, D., … & Trau, M. (2024). Glycan Profiling in Small Extracellular Vesicles with a SERS Microfluidic Biosensor Identifies Early Malignant Development in Lung Cancer. Advanced Science, 2401818.

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