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Stable Isotope Quantitative N-Glycan Analysis by isotope ratio mass spectrometry (CAT#: STEM-ST-0012-LJX)

Introduction

N-glycosylation (a glycan linked to the nitrogen chain of asparagine) is the most widely known form of glycosylation in biologic drugs, especially on Mabs, where n-glycan structure affects pharmacokinetics, pharmacodynamics, and immunogenicity. The effect of glycosylation on protein function means that glycosylation is a key quality attribute (CQA) of the product in biologic drugs, which makes the characterization of N-glycan structure an essential part of the biopharmaceutical development process. Liquid phase separation analysis and subsequent quantitation remains a challenging task for protein-derived oligosaccharides due to their inherent structural complexity and diversity. N-glycan can be quantitatively analyzed by isotope ratio mass spectrometry.




Principle

Isotope ratio mass spectrometry (IRMS) leverages magnetic sector mass spectrometry to enable high-precision measurement of the stable isotope content of a sample. Typical measurements target hydrogen, carbon, nitrogen, and oxygen analyses—although elements with masses up to and including sulfur can be measured. Solid, liquid, or gas phase samples are converted to simple gases then introduced to the IRMS. During analysis, an electron impact source ionizes sample-derived gas which is then accelerated down a flight tube, separated by mass, and quantified using a series of Faraday cups. The high precision of IRMS enables enumeration of even very small isotopic fractionation associated with physical, chemical, and biological transformations or natural abundance measurements.

Applications

For explaining the detailed molecular mechanisms behind biological processes
For understanding and quantifying nutrient and material exchanges between ecosystems
For providing ultra-precise stable isotope analyses
For understanding the geological history of the Earth
For food authenticity, forensic science, medical research and anti-doping testing

Procedure

1. Fill the reaction tube and install it, connect the gas path
2. Check for helium leaks
3. Heat up the reactor, wait for the reaction tube to burn stable, adjust the state of the equipment
4. Wrap the sample in a tin cup and test the sample
5. Store and process data

Materials

• Sample Type:
N-Glycan

Notes

1.The approach is also valuable for quantifying the reactivity and progression of an applied stable isotope tracer to help determine reaction rates and final disposition of applied substrates.
2.IRMS offers a way of measuring isotopic variations with extremely high levels of accuracy. It can be used to detect isotope values of lighter elements with no issues, making it instrumental in the analysis of organic and natural samples.