1. Designing flexible low-viscous sieving media for capillary electrophoresis analysis of ribonucleic acids
- Author
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Gunilla Åkesson Nilsson, Eivor Örnskov, Per-Ola Norrby, Anders Sparén, and Louis De Scheerder
- Subjects
Glycerol ,Resolution (mass spectrometry) ,02 engineering and technology ,01 natural sciences ,Biochemistry ,Analytical Chemistry ,Capillary electrophoresis ,Stability indicating ,medicine ,Prospective Studies ,Chromatography ,Polyvinylpyrrolidone ,Chemistry ,Viscosity ,Design of experiments ,010401 analytical chemistry ,Organic Chemistry ,Electrophoresis, Capillary ,Povidone ,General Medicine ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,Molecular Weight ,High mass ,Separation method ,Drug product ,RNA ,0210 nano-technology ,Gels ,medicine.drug - Abstract
Modified messenger RNA (mRNA) has recently become a new prospective class of drug product. Consequently, stability indicating separation methods are needed to progress pharmaceutical development of mRNA. A promising separation technique for the analysis of mRNA is capillary gel electrophoresis (CGE). We designed a flexible, low-viscous sieving medium for CGE, based on high mass linear polyvinylpyrrolidone (PVP) and glycerol. A Central Composite Face-centered design resulted in a strong model that allowed us to predict suitable sieving media compositions by using multi-objective optimization. The way of working proposed in this paper gives analysts the freedom to design a suitable sieving medium for their response(s) of interest, for purity and stability analysis of polynucleotides with a size around 100–1000 bases. Depending on the criteria for the analysis there will be a trade-off between different suitable conditions. By using this method, we created a sieving medium that was able to improve resolution, peak height and analysis time of an RNA ladder compared to the current commercially available separation gels.
- Published
- 2018