Plasmid purification by using a new naphthalene tripodal support

Santos, Tiago; Proença, Z.; Queiroz, João A.; Tomaz, Cândida T.; Cruz, Carla

doi:10.1016/j.seppur.2017.06.072

Cited by 7 publications

(6 citation statements)

References 25 publications

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“…These values are in good agreement with those previously reported for some of the SILs addressed in this work [ 30 ]. Moreover, based on the nitrogen content provided by the elemental analysis, the ligand density was determined for each modified support [ 35 ], being given in Table 1 . These values are useful to address the tRNA binding ability and nucleic acids separation performance (addressed below).…”

Section: Resultsmentioning

confidence: 99%

Efficient Isolation of Bacterial RNAs Using Silica-Based Materials Modified with Ionic Liquids

Pereira

Pedro

Neves

et al. 2021

Life

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High quality nucleic acids (with high integrity, purity, and biological activity) have become indispensable products of modern society, both in molecular diagnosis and to be used as biopharmaceuticals. As the current methods available for the extraction and purification of nucleic acids are laborious, time-consuming, and usually rely on the use of hazardous chemicals, there is an unmet need towards the development of more sustainable and cost-effective technologies for nucleic acids purification. Accordingly, this study addresses the preparation and evaluation of silica-based materials chemically modified with chloride-based ionic liquids (supported ionic liquids, SILs) as potential materials to effectively isolate RNAs. The investigated chloride-based SILs comprise the following cations: 1-methyl-3-propylimidazolium, triethylpropylammonium, dimethylbutylpropylammonium, and trioctylpropylammonium. All SILs were synthesized by us and characterized by solid-state 13C Nuclear Magnetic Resonance (NMR), Scanning Electron Microscopy (SEM), elemental analysis, and zeta potential measurements, confirming the successful covalent attachment of each IL cation with no relevant changes in the morphology of materials. Their innovative application as chromatographic supports for the isolation of recombinant RNA was then evaluated. Adsorption kinetics of transfer RNA (tRNA) on the modified silica-based materials were investigated at 25 °C. Irrespective to the immobilized IL, the adsorption experimental data are better described by a pseudo first-order model, and maximum tRNA binding capacities of circa 16 µmol of tRNA/g of material were achieved with silica modified with 1-methyl-3-propylimidazolium chloride and dimethylbutylpropylammonium chloride. Furthermore, the multimodal character displayed by SILs was explored towards the purification of tRNA from Escherichia coli lysates, which in addition to tRNA contain ribosomal RNA and genomic DNA. The best performance on the tRNA isolation was achieved with SILs comprising 1-methyl-3-propylimidazolium chloride and dimethylbutylpropylammonium chloride. Overall, the IL modified silica-based materials represent a more efficient, sustainable, and cost-effective technology for the purification of bacterial RNAs, paving the way for their use in the purification of distinct biomolecules or nucleic acids from other sources.

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Section: Resultsmentioning

confidence: 99%

Efficient Isolation of Bacterial RNAs Using Silica-Based Materials Modified with Ionic Liquids

Pereira

Pedro

Neves

et al. 2021

Life

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“…The elemental analysis results given in Table 1 show no significant differences in the carbon and hydrogen content ( wt %) for both matrices, namely MP and MP-SilPrMImCl supports; however, nitrogen (2.03 wt %) was detected in the final modified support, as well as in MP-MIm support, confirming the presence of the imidazolium ring. Based on the nitrogen content provided by elemental analysis, the ligand density [ 41 ] of MP-SilPrMImCl (given in Table 1 ) corresponds to 0.725 mmol of IL per gram of support, a higher value than that present in supports modified with common ligands used in the separation of nucleic acids (naphthalene tripodal: 0.32 mmol/g sepharose CL-6B [ 41 ], and 3,8-diamino-6-phenylphenathridine: 0.15 mmol/g derivatized sepharose [ 42 ]). Taking all characterization results into consideration, it is shown that the IL is efficiently and covalently bound to the support.…”

Section: Resultsmentioning

confidence: 99%

Improved ionic-liquid-functionalized macroporous supports able to purify nucleic acids in one step

Neves

Pereira

Pedro

et al. 2020

Materials Today Bio

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“…Recent chromatographic procedures related to purification of pDNA vaccines include anion exchange chromatography, affinity chromatography, multimodal chromatography, size exclusion and hydrophobic interaction chromatography [ 97 , 99 , 100 , 101 , 102 ].…”

Section: Analytical Approachesmentioning

confidence: 99%

mRNA Therapeutic Modalities Design, Formulation and Manufacturing under Pharma 4.0 Principles

et al. 2021

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In the quest for a formidable weapon against the SARS-CoV-2 pandemic, mRNA therapeutics have stolen the spotlight. mRNA vaccines are a prime example of the benefits of mRNA approaches towards a broad array of clinical entities and druggable targets. Amongst these benefits is the rapid cycle “from design to production” of an mRNA product compared to their peptide counterparts, the mutability of the production line should another target be chosen, the side-stepping of safety issues posed by DNA therapeutics being permanently integrated into the transfected cell’s genome and the controlled precision over the translated peptides. Furthermore, mRNA applications are versatile: apart from vaccines it can be used as a replacement therapy, even to create chimeric antigen receptor T-cells or reprogram somatic cells. Still, the sudden global demand for mRNA has highlighted the shortcomings in its industrial production as well as its formulation, efficacy and applicability. Continuous, smart mRNA manufacturing 4.0 technologies have been recently proposed to address such challenges. In this work, we examine the lab and upscaled production of mRNA therapeutics, the mRNA modifications proposed that increase its efficacy and lower its immunogenicity, the vectors available for delivery and the stability considerations concerning long-term storage.

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Plasmid purification by using a new naphthalene tripodal support

Cited by 7 publications

References 25 publications

Efficient Isolation of Bacterial RNAs Using Silica-Based Materials Modified with Ionic Liquids

Efficient Isolation of Bacterial RNAs Using Silica-Based Materials Modified with Ionic Liquids

Improved ionic-liquid-functionalized macroporous supports able to purify nucleic acids in one step

mRNA Therapeutic Modalities Design, Formulation and Manufacturing under Pharma 4.0 Principles

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