Ribonucleic acid purification

Current progress on microRNAs‐based therapeutics in neurodegenerative diseases

“…While this method is able to produce RNA molecules on a microgram scale, large‐scale productions are usually hampered by the need for high quantities of expensive RNA polymerases . In general, although these methods are efficient to miRNA production, additional isolation and purification protocols are required in order to remove process‐associated impurities (linearized plasmid DNA template, enzymes, unreacted nucleotides, salts or buffers, short aberrant transcripts, failure in the sequences, among others) . Indeed, the final RNA products can be purified using preparative polyacrylamide gel electrophoresis followed by gel extraction, size exclusion chromatography, and anion exchange chromatography (see subsection MicroRNA Purification ) .…”

“…In turn, anion exchange chromatography relies not only on the polyanionic nature of RNA molecules but also on their molecular size . Although these methods can be very efficient for the purification of RNA under nondenaturing conditions, at low to moderate operating pressures, some of these still require several time‐consuming preparatory steps (such as phenol/chloroform extractions to remove proteins followed by desalting and sample concentration) and are expensive on large scale . For these reasons, the RNA product purified by these strategies may require additional treatments to be suitable for use in analytical and clinical applications, and therefore it may be difficult to maintain the stability and biological activity of the target RNA.…”

“…In the last years, the downstream strategies based on affinity chromatography have been developed in an attempt to circumvent several challenges in RNA purification, and actually they represent a highly efficient option for RNA purification to increase not only the yields but also the selectivity in a single chromatographic step . Most approaches described in affinity chromatography make use of RNA affinity tags (small sequences that are introduced in the RNA molecules) that bind with high affinity to specific molecules, used as ligands in chromatographic matrices for the selective purification of RNAs from cellular extracts .…”

Pharmaceutical-grade pre-miR-29 purification using an agmatine monolithic support

“…The purification of RNA molecules is already reported by using preparative denaturing polyacrylamide gel electrophoresis (PAGE), http://dx.doi.org/10.1016/j.chroma.2014.09.080 0021-9673/© 2014 Elsevier B.V. All rights reserved. affinity tag-based purification, anion-exchange and size exclusion chromatography [7][8][9][10][11]. These methods make use of enzymes and structural modifications in the RNA molecule by the introduction of tags sequences, which can affect the overall topology of the RNA.…”

New insights for therapeutic recombinant human miRNAs heterologous production: Rhodovolum sulfidophilum vs Escherichia coli

“…Actually, this fact still restricts the application of these RNAs onto preclinical or clinical trials. 15,16 In particular, beyond the fact that large-scale chemical synthesis of RNA remains technically difficult and expensive, these RNAs are traditionally purified by preparative electrophoresis followed by gel extraction, a procedure that is time-consuming and leads to acrylamide-associated impurities within the RNA. 17 As an alternative to overcome these limitations, huge efforts have been made in recent years to develop recombinant RNA techniques to cost-effectively produce biologic RNA agents in vivo that can better retain the structure, function and safety properties of natural RNAs.…”