In addition, single stranded DNA often allows for a free exposure of the hydrophobic aromatic bases. These include a negatively charged phosphate backbone as well as a hydrophobic character originating mainly from the major groove of DNA which exposes the base pairs on the surface of the molecule. DNA molecules harbour some intrinsic chemical properties that render them suitable for chromatographic separations. The needs for purified nucleic acids for preparative and analytical applications have increased constantly, demanding for the development of new and more efficient methods for their recovery and isolation. Gel electrophoresis showed high assembly yield and purity, whereas fluorescence correlation spectroscopy confirmed that the tetrahedrons had a diffusion coefficient (26.7 μm² s⁻¹) consistent with the expected size (20 nm). In either case, collected ssDNA-containing fractions were homogeneous and impurity free.įinally, 8.4 μg of a 1000-nt ssDNA fragment was purified and used alongside with site-specific short oligonucleotides (staples) to assemble 63-bp edge length tetrahedrons. In multimodal chromatography, however, the elution pattern was reversed, highlighting the importance played by hydrophobicity. In anion exchange chromatography, the less-charged ssDNA eluted before the dsDNA. To isolate the target ssDNA from dsDNA and other PCR impurities, anion-exchange (Q-ligand) and multimodal chromatography (CaptoTM adhere ImpRes) were explored using stepwise gradients with increasing NaCl concentrations. Alternatively, we present a chromatography-based method to purify ssDNA scaffolds from aPCR mixtures, which can be used in the context of DNA-origami techniques.ĪPCR was performed to generate single and double-stranded DNA (dsDNA) from the M13mp18 genome. Each scaffold is usually purified by agarose gel extraction, a technique that is laborious, limited, not scalable, presents low recovery yields and a low-quality product. When working with sensitive MAbs, the wide window of operation simplifies optimization of conditions.Ĭapto MMC ImpRes is available in a range of different bulk pack sizes for easy scale-up and pre-packed formats for process development.Īs member of the BioProcess media range, Capto MMC ImpRes meets industrial demands with security of supply and comprehensive technical and regulatory support.DNA-origami biomanufacturing relies in many cases on the use of asymmetric PCR (aPCR) to generate 500-3500 base, object-specific, single-stranded DNA (ssDNA) scaffolds. The multimodal cation exchanger ligand of Capto MMC ImpRes is immobilized to the base matrix and interacts with the target molecule through multiple types of interactions.Ĭapto MMC ImpRes can be used for initial or final polishing steps in MAb purification processes, as well as for polishing of antibody fragments such as domain antibodies (DAbs).The wide window of operation for conductivity and/or pH simplifies the process as conditioning after the protein A step can be omitted. The weak cation exchange multimodal ligand enables high selectivity in a broad pH/salt window compared with traditional ion exchangers, which allows the use of Capto MMC ImpRes in a variety of process conditions to solve challenging purification problems. High yields achieved through the high-resolution beads and selectivity of the ligand.Efficient removal of aggregates, viruses, and main contaminants in MAb processes.Enables use of a platform approach to MAb process development.Displays a broad pH/salt operational window.Suitable for polishing of antibody fragments.Capto MMC ImpRes is a weak cation exchange multimodal BioProcess chromatography medium (resin), designed to meet the demands of modern large-scale manufacturers, for high-resolution polishing of monoclonal antibodies (MAbs) and other biomolecules. Suitable for polishing of antibody fragments.Displays a broad pH/salt operational window.
Capto MMC ImpRes is a weak cation exchange multimodal BioProcess chromatography medium (resin), designed to meet the demands of modern large-scale manufacturers, for high-resolution polishing of monoclonal antibodies (MAbs) and other biomolecules.
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