Hi,
We want to perform glycan analysis using tissue samples (human and mouse). We consider to purchase a capillary electrophoresis machine P/ACE MDQ, however we are not sure if it is user friendly. Personally I do not have much experience with biochemical approaches, I would like to hear your opinion if someone without experience can easily run the machine and how much of data we can get from CE-LIF (especially compared to mass spec).
Thank you.
Hi there -
I don't want to make a recommendation for a specific vendor, but for the questions you have regarding user friendliness of CE and comparison with MS, I can make a few comments.
"User friendliness" is relative of course, but for someone with knowledge of basic biochemical protocols for sample prep etc should be able to attempt running it. While CE is not exactly a push-button approach, it is not too difficult. The instrument makers provide pretty good training. For CE-LIF, you can check out their webinar. Finally, comparison with MS data is challenging. MS can be applied to conduct a wide variety of research on glycans including glycan distribution, site of attachment etc. CE-LIF gives you distribution of released glycans from the glycoprotein of interest. Although they are not apple-to-apple comparison, glycan profile should be similar in both (CE, MS) with CE often providing more quantitative data on glycan distribution.
At the end, you need to conduct good due diligence before making the significant investment. Hope it helps.
Please look at the following below links which may help you in your analysis:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6135675/pdf/cr7b00669.pdf
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2898890/pdf/nihms-216552.pdf
https://sciex.com/Documents/tech%20notes/products/ce/single-analytical-platform-for-glycan-analysis-charge-heterogeneity.pdf
Thanks
Excellent advice from Das and very good references from Elgailani.
If you use good laboratory practices, you should not have too much trouble with capillary electrophoresis. My first system used two microfuge tubes, some copper wiring and a scavenged power supply. I did not even have a detector but measured dyes by eye and stopwatch. I also learned that it is easy to block a capillary and that all buffers should be vacuum filtered through 0.2 micron filter..
As for glycan analysis--what do you want to do? If you are just looking for a profile, you can get great results by CE. But if you want to determine the structure of previously unidentified glycans, then it is hard to get enough material. MS might be better, in that case. Or switching totally to something like HPAE-PAD so you can collect the peak and do follow-up experiments.
I will assume that you are more of a biologist and are not looking at the glycans for their own interesting chemistry. In that case, as the first paper from Elgailani shows that you can get phenomenal separations via CE. And, since the chemistry of adding the fluorophore is virtually quantitative, you can get good results on the amounts of glycans present. And there are many more papers on using CE for glycan analysis. You may want to check out the website of the Complex Carbohydrate Research Center ( https://www.ccrc.uga.edu/ ) for more information.