Better capture options are needed to meet the needs of large clinical trials, scale-up, and manufacture of vaccines. Hydrophobic interaction chromatography (HIC) and reverse phase chromatography (RPC) separate ssRNA from dsRNA and short transcripts, but their sensitivity to fouling by proteins and aggregates makes them better suited for polishing than for capture. Traditional anion exchangers elute only mRNA smaller than about 500 bases unless the columns are heated to 50–70☌. It can be used for capture but it cannot discriminate dsRNA (double-stranded) from ssRNA (single-stranded) and it supports only brief cleaning with 100 mM sodium hydroxide. Hybridization-affinity uses a polythymidine (Oligo dT) ligand to base-pair with the polyadenine tail of mRNA. One of the barriers to development of industrial purification platforms for large mRNA has been an inadequate selection of high-performing capture-purification tools. Pete Gagnon, Blaž Goričar, Špela Peršič, Urh Černigoj, Aleš Štrancar
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