NEB TV Ep. 17 – Glycobiology and Clinical Applications

Learn about glycobiology and its importance in clinical and diagnostic applications in this episode of NEB TV. Also, hear more about how NEB is setting the bar for product quality in this rapidly growing field.

Script

NEB TV, what's trending in science.

 

Deana Martin:

Welcome to NEB TV. Today, I'm joined by Alicia Bielik who is a group leader on our production team.  Hi Alicia.

 

Alicia Bielik:

Hi Deana.

 

Deana Martin:

What are we talking about today?

 

Alicia Bielik:

Today, I'd like to introduce you to the importance of glycobiology. Then, you will hear from two experts in the field, Pauline Rudd and Gordan Lauc, on the importance of clinical and diagnostic glycoscience, as well as pharmaceutical glycoscience. Stephen Shi and I will then describe the breath of NEB's glycobiology product line and our commitment to the highest quality standards.

 

Deana Martin:

Great! Are you ready?

 

Alicia Bielik:

I'm ready.

 

Deana Martin:

Let's get started.

 

Science in 60

 

Alicia Bielik:

Glycobiology is the study of the structure, function, and biology of carbohydrates, which are also called glycans, and are present in every living organism. It's a rapidly growing field in biology with relevance to biomedicine, biotechnology, and basic research.

 

Proteomics has expanded the knowledge of protein expression, modification, interaction, and function.

 

In eukaryotic cells, the majority of proteins are post-translationally modified. One of the most common modifications, which is essential for cell viability, is the attachment of glycans, which is also known as glycosylation. These glycans are diverse complex molecules whose function is determined by their unique structure.

 

Effective enzymatic tools and analytical techniques are required to properly elucidate glycan structure, which then can be correlated to a specific function.

 

Glycosylation defines the adhesive properties of proteins and cells. In fact, the immune system largely functions via glycan protein interactions. This highlights the importance of glycans in physiology, pathogen recognition, cancer, and autoimmune diseases.

 

Considering the essential role that glycans play in immune interactions, glycoproteins and carbohydrates are becoming the next generation of therapeutics, vaccines and diagnostics.

 

Clinical and Diagnostic Glycoscience Applications

 

Why are glycans important in pharmaceutical chemistry?

 

Pauline Rudd:

Most of the new biological drugs are glycosylated proteins. The glycans have a number of fairly important functions in the roles of those proteins. So, for example in Erythropoietin, which is probably the biggest blockbuster drug at the moment, needs the correct sugars attached to the proteins in order to maintain a useful half-life for the protein. If you have the wrong sugars, it will be cleared in three minutes. If you have the right ones, it will be in the patient for at least three hours, in which case it has time to act in its biological role.

 

Monoclonal antibodies also have sugars which modulate the effective function of the proteins. So they're generally targeted to epitopes on cancer cells for example. When they engage with those epitopes it's really important to control the next stage of the process which is for the Fc region of the molecule to bind to some other receptor and then to activate a biological event such as antigen dependence and cellular cytotoxicity.

 

We also know that glycans can be pro- or anti-inflammatory molecules. So all these drugs are designed to act with the patient's immune system. If for example, the IgG molecules have sugars ending in salicylic acids, then these are basically anti-inflammatory molecules. So it's very important to have an understanding of these different roles and functions of the sugars before you design the kind of antibody glycosylation that you need for your particular purpose.

 

Are glycans promising molecules for monitoring health and/or disease?

 

Gordon Lauc:

Glycans change both with healthy aging and with disease. In the last few years in GENOS we have analyzed over 50,000 individual glycans. In the majority of studies we found significant changes.

 

In many diseases that we've studied, including diabetes, cancer, and IVT( Idiopathic ventricular tachycardia), changes in the composition of either total plasma glycome or the IgG glycome contained more information about the disease than all other clinical parameters combined.

 

Therefore, glycan profiling is definitely a very promising biomarker.

 

Why are the glycans of biologic drugs structurally profiled?

 

Pauline Rudd:

Well it's very important to profile and to have a detailed analysis of the sugars attached to your drug. For example you need to be certain that there are no antigenic epitopes because sometimes if you make a drug in a non-human cell line, you may end up with a epitope on the sugar which will react with the immune system and will cause a very severe response. Since that has happened a number of times, the regulators really press home the importance of checking for this. It's also important to monitor the glycosylation because it is the end point of any production of a glycoprotein. If it's consistent, it's telling you that the bioprocess itself is consistent.

 

Where do you see clinical glycomics in the next five years?

 

Gordon Lauc:

Clinical glycomics is clearly heading towards a new revolution. Analytical methods are improving a lot and nowadays they enable a routine high throughput analysis of both total plasma glycome and some individual glycoproteins.

 

This was very clearly demonstrated in a recent large collaborative study that was performed by GENOS, LEMC, NIBRT, and NPI. The availability of technology in the first large clinical studies that were published in the last few years, are also raising interest among clinical colleagues who are showing higher and higher interest in glycans in their studies.

 

The field of personalized medicine is surely one big area where glycomics is expanding. Diagnosing and monitoring diseases is currently the focus of research, but even more promising seems to be the development of companion diagnostics that will help predict the response to different therapeutics. Since glycan is both highly heritable and highly versatile at the same time, it seems to be an ideal tool for stable patients certification, which is exactly what we is needed for precision medicine.

 

Where is glycan analysis headed in the next five years?

 

Pauline Rudd:

Our focus now is on further miniaturization. We would like to see it transferred onto a chip for example so that it could easily fit in at line in a manufacturing process. We'd also like to see the analytics being quite faster than the current LCMS system. I think those are two major targets. The high throughput robot, which we use to do multiple releases simultaneously, is also going to be very important because people are now moving towards establishing a human glycome project which will be looking at the glycosylation of serum or individual molecules from many thousands of people.

 

Setting the Bar For Product Quality

 

Stephan Shi:

As you have heard from leading experts, glycosylation is increasing in its importance to regulated industries like pharma and clinical diagnostics. Many glucosidase manufacturers are still using practices designed to support the research market.

 

Recently, we tested analytical glycosidases from several commercial providers for various quality attributes. We found a range of issues that highlight the need for revised manufacturing practices for glycosidases used in pharma and clinical applications. For example one alpha-glucosidase of the glycosidases from one supplier produced from coffee bean turned out to have substantial beta-glycosidase activity.  In another case, two of the glycosidase from different suppliers we examined were found to contain sialidase activity.

 

The presence of unwanted enzyme activities can obviously taint experimental results especially when glycosidases are used alone or while in combinations to elucidate or sequence elaborate glycan structures.

 

Alicia Bielik:

The quality of NEB's products is both an NEB strength and a differentiator from other providers. It's our fundamental promise to both the scientific community and every NEB customer.

 

We have a comprehensive quality management system that insures every step in the manufacturing process is held to the highest quality standards in the business.  One indication is our certification to ISO 9001 quality management system as well as ISO 13485, which is a requirement for the medical device industry.

 

NEB is also building a GMP facility, which when complete will elevate our quality standards for certain products even higher.

 

We have created set of stringent quality control standards for every enzyme that we produce. Including a protein purity standard of >%95 pure and a sensitivity assay that includes a panel of 18 substrates for the detection of contaminating exo- or endo-glycosidase activities. Each enzyme is also tested for the absence of protease activity.

 

In this way we are able to manufacture enzymes with unmatched quality standards and exceptional lot to lot consistency.

 

Deana Martin:

So Alicia, thanks so much for joining us today.

 

Alicia Bielik:

Thanks for having me.

 

Deana Martin:

As always, if you have any suggestions for future episodes, please let us know.

 

"GMP-grade" is a branding term NEB uses to describe reagents manufactured at NEB’s Rowley facility. The Rowley facility was designed to manufacture reagents under more rigorous infrastructure and process controls to achieve more stringent product specifications and customer requirements. Reagents manufactured at NEB’s Rowley facility are manufactured in compliance with ISO 9001 and ISO 13485 quality management system standards. However, at this time, NEB does not manufacture or sell products known as Active Pharmaceutical Ingredients (APIs), nor does NEB manufacture its products in compliance with all of the Current Good Manufacturing Practice regulations. 

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