Filter: Nucleic acids
2020

CIMac PrimaS™ is a new member of BIA’s family of high performance monoliths for analysis and purification of mRNA. Its positive charge gives it some anion exchange behavior but hydrogen bonding makes its selectivity is entirely distinct from traditional anion exchangers. QA and DEAE anion exchangers need to be heated into the range of 50–70°C for large mRNA to elute. PrimaS elutes mRNA in a pH gradient, well separated from DNA and dsRNA. New experimental data show that NaCl shifts the elution profile to lower pH values.

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CIMac PrimaS is a new member of BIA’s family of high performance chromatography products for analysis and purification of mRNA. Its positive charge gives it some anion exchange behavior but hydrogen bonding makes its selectivity is entirely distinct from traditional anion exchangers. QA and DEAE anion exchangers need to be heated into the range of 50–70°C for large mRNA to elute. Large mRNA elutes from CIMac PrimaS at ambient temperature in an ascending pH gradient. New experimental data show that large mRNA can also be eluted from CIMac PrimaS at ambient temperature in a pyrophosphate gradient. Resolution between in vitro transcription (IVT) contaminants and mRNA can be controlled with pH.

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CIMmultus PrimaS purifies large single-stranded mRNA (ssRNA) under aqueous conditions at ambient temperature. It removes dsRNA, DNA, proteins, and endotoxins while fractionating ssRNA in order of increasing size. It can be used for one-step purification of research grade ssRNA or as a high-resolution capture step in a multi-step purification process. It also enables fast high-resolution analytical characterization of in vitro transcription mixtures, partially purified samples, chromatography fractions, and formulated drug substance.

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CIMmultus C4 HLD is a hydrophobic interaction chromatography monolith for purification of nucleic acids. Sample is applied at high ionic strength in salts that precipitate RNA. The column is eluted with a decreasing salt gradient. The majority of DNA, dsRNA, and short transcripts elute earlier than intact ssRNA. The majority of proteins and aggregates bind very strongly and are eliminated by a cleaning step with NaOH. CIMmultus C4 HLD can be used to produce research grade ssRNA from in vitro transcription (IVT) mixtures but gives its best results as a polishing method.

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CIMmultus Oligo dT is a hybridization-affinity chromatography monolith for purification of mRNA that terminates in a poly-A tail. Non-RNA contaminants flow through the column during sample loading. RNA with a poly-A terminus binds. The column is washed then the mRNA is recovered in a single elution step at neutral pH. CIMmultus Oligo dT can be used for one-step purification of research grade ssRNA, as a high-resolution capture method in a multi-step purification process, as a polishing method, or as an analytical method for estimating quantity and purity of mRNA in a sample. It can be used with mRNA up to 10 kb or more.

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