“…The recovery of the small RNA from the IL-functionalized matrix in the binding step is highly advantageous because it elutes with a low salt concentration, as well as with a low residence time within the chromatographic system—two crucial factors for maintaining the RNA stability given its high susceptibility to degradation by ribonucleases. It should be remarked that these factors are not always obeyed in conventional strategies to separate RNA, such as lysine affinity chromatography [ 47 ] and agmatine affinity chromatography [ 48 ] where small RNA is recovered with aqueous solutions of (NH 4 ) 2 SO 4 at 1.5 M and NaCl at 1.75 M, respectively.…”
Section: Resultsmentioning
confidence: 99%
“…The MP-SilPrMImCl DBC for BSA, plasmid DNA, and small RNA at 10% of maximum capacity is, respectively, 37.0, 0.60, and 3.68 mg/mL, while at 50% of maximum capacity it is, respectively, 59.1, 0.83, and 12.0 mg/mL. The values of the MP-SilPrMImCl binding capacity toward RNA are particularly remarkable, especially when compared to the agmatine-based monolith, where 0.71 and 1.37 mg/mL of DBC, at 10% and 50% of maximum capacity, respectively, have been reported [ 48 ], the HPLC 21.2 mm × 100 mm RNASep™ Semi-Prep Cartridge (ADS Biotec, Omaha, USA) wherein a load capacity of 600 μg total RNA is achievable, or the loading capacities ranging from 0.6 to 8.0 mg double stranded RNA/mL of resin using distinct anion-exchange columns [ 52 ]. Fig.…”
“…The recovery of the small RNA from the IL-functionalized matrix in the binding step is highly advantageous because it elutes with a low salt concentration, as well as with a low residence time within the chromatographic system—two crucial factors for maintaining the RNA stability given its high susceptibility to degradation by ribonucleases. It should be remarked that these factors are not always obeyed in conventional strategies to separate RNA, such as lysine affinity chromatography [ 47 ] and agmatine affinity chromatography [ 48 ] where small RNA is recovered with aqueous solutions of (NH 4 ) 2 SO 4 at 1.5 M and NaCl at 1.75 M, respectively.…”
Section: Resultsmentioning
confidence: 99%
“…The MP-SilPrMImCl DBC for BSA, plasmid DNA, and small RNA at 10% of maximum capacity is, respectively, 37.0, 0.60, and 3.68 mg/mL, while at 50% of maximum capacity it is, respectively, 59.1, 0.83, and 12.0 mg/mL. The values of the MP-SilPrMImCl binding capacity toward RNA are particularly remarkable, especially when compared to the agmatine-based monolith, where 0.71 and 1.37 mg/mL of DBC, at 10% and 50% of maximum capacity, respectively, have been reported [ 48 ], the HPLC 21.2 mm × 100 mm RNASep™ Semi-Prep Cartridge (ADS Biotec, Omaha, USA) wherein a load capacity of 600 μg total RNA is achievable, or the loading capacities ranging from 0.6 to 8.0 mg double stranded RNA/mL of resin using distinct anion-exchange columns [ 52 ]. Fig.…”
“…Many monoliths that have been used in affinity chromatography are polymers based on glycidyl methacrylate (GMA) and ethylene glycol dimethacrylate (EDMA), which have been used with immobilized agents that include antibodies, enzymes, and peptides [42,58,[61][62][63]. This includes polymethacrylate monoliths known as convective interaction media (CIM), which have been employed in applications that range from the purification of large biological agents (e.g., DNA, proteins, and viruses) [60] to antibody-based separations [64,65] and immobilized metal-ion affinity chromatography (IMAC) [66][67][68][69].…”
Section: Supports In Affinity Chromatographymentioning
confidence: 99%
“…As shown in Figure 6, metal ion chelates are another important example of non-biological binding agents that can be used in affinity chromatography. This combination is the basis of the method of IMAC [66][67][68][69], which was briefly introduced in Section 4. In 1975, Porath et.…”
“…These ligands simulate and exploit naturally occurring biological interactions, combining multiple noncovalent interactions, that occur at the cellular level . The RNA structural features (such as negative charge of the RNA backbone, structural diversity of RNAs, RNA conformational rearrangement, base composition, and high base exposure on RNA species) seem to be relevant for the specific interactions established between the immobilized ligands (amino acids and derivatives, namely O ‐ phospho‐ l ‐tyrosine, l ‐arginine, l ‐lysine, and Agmatine) and the target pre‐miRNAs. Globally, the use of amino acids immobilized in bead‐packed columns or monolithic columns allowed to recover and separate the target pre‐miRNA from complex extracts.…”
Section: Micrornas As Therapeutic Productsmentioning
MicroRNAs (miRNAs)-based therapy has recently emerged as a promising strategy in the treatments of neurodegenerative diseases. Thus, in this review, the most recent and important challenges and advances on the development of miRNA therapeutics for brain targeting are discussed. In particular, this review highlights current knowledge and progress in the field of manufacturing, recovery, isolation, purification, and analysis of these therapeutic oligonucleotides. Finally, the available miRNA delivery systems are reviewed and an analysis is presented in what concerns to the current challenges that have to be addressed to ensure their specificity and efficacy. Overall, it is intended to provide a perspective on the future of miRNA-based therapeutics, focusing the biotechnological approach to obtain miRNAs. WIREs RNA 2017, 8:e1409. doi: 10.1002/wrna.1409 For further resources related to this article, please visit the WIREs website.
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