iBB
46.4K views | +5 today
Follow
iBB
Institute for Bioengineering and Biosciences
Curated by iBB
Your new post is loading...
Your new post is loading...
Scoop.it!

Magnetic Particles for the Purification of DNA Scaffolds for Biomanufacturing DNA-Origami Nanostructures

Magnetic Particles for the Purification of DNA Scaffolds for Biomanufacturing DNA-Origami Nanostructures | iBB | Scoop.it

Asymmetric PCR (aPCR) is often used to generate single-stranded DNA (ssDNA) scaffolds, which can then be assembled into nanoobjects by the DNA-origami technique. The scaffolds are usually purified by agarose gel extraction, a laborious, time consuming, limited, and non-scalable technique that presents low recovery yields, delivers low-quality products. To overcome such pitfalls, Ana Silva-Santos, Rui Silva, Sara Rosa and Miguel Prazeres from BERG-iBB, in collaboration with Pedro Paulo from CQE developed a simple, fast, and potentially scalable affinity-based method comprising magnetic particles. Specifically, scaffolds were synthesized by aPCR and purifed using magnetic particles functionalized with a 20 nt oligonucleotide complementary to the 3′ end of the target. The purified scaffolds were used to assemble 31 and 63 bp edge length tetrahedra with short oligonucleotides and thermal annealing, demonstrating the potential of affinity-based magnetic beads in the production of DNA-origami nanostructures. The work was published in ACS Applied Nanomaterials.

No comment yet.
Scoop.it!

Featured Photo: Illustration of DNA Origami Nanotetrahedron

Featured Photo: Illustration of DNA Origami Nanotetrahedron | iBB | Scoop.it

Description: Illustration of a DNA origami nanotetrahedron created with David Goodsell’s Illustrate software.

 

Context: BIOTecnico PhD student Ana Rita Santos, under the supervision of BERG-iBB researcher Miguel Prazeres is developing processes to produce and purify DNA scaffolds and DNA origami nano-objects.

No comment yet.
Scoop.it!

Purification of Plasmid DNA by Multimodal Chromatography

Purification of Plasmid DNA by Multimodal Chromatography | iBB | Scoop.it
Production of plasmid DNA is important in the context of gene therapy and DNA vaccination. In a paper just published in the journal Separation and Purification Technology, Ana Rita Santos, Cláudia Alves, Ana Azevedo and Miguel Prazeres from BERG-iBB, describe a process based on tandem precipiatation and multimodal chromatography that yields pure supercoiled plasmid DNA. The chromatography step, which uses a cationic multimodal ligand (Capto™ adhere) and an NaCl step gradient, provides baseline separation of isoforms and yields supercoiled-rich fractions (>90%) that are virtually free from RNA and have levels of gDNA and protein impurities within specifications. Click on title to learn more.
No comment yet.
Scoop.it!

BERG Research Presented at SBCN 2021

BERG Research Presented at SBCN 2021 | iBB | Scoop.it

The 15th international symposium on Biochromatography and Nanoseparations, which is taking place from 17-20th May in virtual mode, aims to highlight the recent progress on the purification and characterization of biomolecules from lab to industrial scale. BERG-IBB researchers will be presenting their work with oral (Ana Azevedo, Ana Rita Santos, Amin Javidanbardan and Malik Wahab) and poster (Jorge João) communications.

No comment yet.
Scoop.it!

Multimodal Chromatography of Supercoiled Minicircles

Multimodal Chromatography of Supercoiled Minicircles | iBB | Scoop.it

The ability to purify supercoiled minicircles (sc MC) from related miniplasmid (MP) and parental plasmid (PP) impurities is critical for a wider application of MC vectors in gene therapy research. BERG-iBB researchers developed a process that relies on multimodal chromatography with a 2-step NaCl elution to separate sc MC from open circular impurities. A detailed study of the DNA-ligand interactions underlying the separation shows that the order of elution reflects the increasing degree of base exposure of the species. Data further suggests that nucleic acid bases most likely engage in cation-π, π-π stacking and H-bonding with ligand. The work was published in Separation and Purification Technology.

No comment yet.
Scoop.it!

Separation of Plasmid DNA Topoisomers by Multimodal Chromatography

Separation of Plasmid DNA Topoisomers by Multimodal Chromatography | iBB | Scoop.it
The ability to analyze the distribution of topoisomers in a plasmid DNA sample is important when evaluating the quality of preparations intended for gene therapy and DNA vaccination, or when performing biochemical studies on the action of topoisomerases and gyrases. In a recent paper in Analytical Biochemistry, BERG-iBB researchers led by Miguel Prazeres and Ana Azevedo we describe the separation of supercoiled (sc) and open circular (oc) topoisomers by multimodal chromatography. A medium modified with the ligand N-benzyl-N-methyl ethanolamine and an elution scheme with increasing NaCl concentration are used to accomplish the baseline separation of sc and oc plasmid. Click on title to learn more.
No comment yet.