Webinar Review: Making Sense of Glycosylation Using Novel Hydrophilic Interaction Liquid Chromatography Techniques

Our trainees review webinars in their given fields and share abstracts to help colleagues outside their discipline make an informed choice about watching them. As our program bridges diverse disciplines, these abstracts are beneficial for our own group in helping one another gain key knowledge in each other’s fields. We are happy to share these here for anyone else who may find them helpful.

Making Sense of Glycosylation Using Novel Hydrophilic Interaction Liquid Chromatography Techniques

Matthew Lauber, Ph.D
Principal Applications Chemist (Waters Corporation)

November 17, 2015 at 11 AM ET

Waters Corporation

Watch on the Waters website >>

Shannon BernecheAnalysis by Shannon Berneche:

This webinar provides an overview of a technique called hydrophilic interaction liquid chromatography (known as HILIC), which I am using in my external research experience lab for separating glycoproteins to allow their identification (which can be done via mass spectrometry as well as other detection methods following their separation by HILIC). This is also the application being described in this webinar, and it is an interesting topic because glycoproteins are very important in the body as signaling molecules and other functions, and their abnormal expression likely plays a role in many diseases, but they are not yet very well characterized.

Glycoprotein profiling is an area in the field of mass spectrometry which is still under development but can potentially be very valuable once it is optimized, as it can then be applied to understand the mechanism of various disease states which involve dysregulation of these important molecules. Glycoproteins also show potential to be used as possible drugs to treat many different diseases, making it important to be able to analyze them in a standardized way.

In this webinar, the speaker first covers the basics of how HILIC works. It’s important to understand this in order to understand how one would be able to apply this in their research, as well as the results obtained through this method. He then explains why HILIC is useful for the analysis of glycoproteins, which face unique challenges compared to analyzing other types of molecules with liquid chromatography. There are several reasons, as he explains, that HILIC uniquely aids in analyzing these molecules and why it is needed for this particular application (and some others) as opposed to other widely used types of liquid chromatography.

He also explains that there are many different ways to analyze glycans (bottom-up, top-down, middle-down, intact or denatured) which are important to consider in your applications and in understanding different types of experiments. Another important consideration is the sample preparation workflow which can be used before HILIC, for which there are many options. Although he focuses on specific products which this company is selling, it is important to find the one that is optimized for your applications, and it is useful to see examples of products that you may be able to use in your sample prep. Finally, data processing is necessary to consider for this method, in order to interpret the data in the most effective way possible and to understand your results. He explains various techniques that can be used for each of these components of HILIC analysis of glycoproteins, all of which are very important for scientists who plan to use this technique in their research.

Overall, this webinar is very useful for those who wish to implement HILIC in their own research, and will also be interesting to those who would like to learn more about how glycoproteins can be identified. If you are interested in learning about methods for glycoprotein analysis, I would recommend watching this webinar.