![]() doi: 10.1016/j.chroma.2004.03.022Įon-Duval A, Burke G (2004) Purification of pharmaceutical-grade plasmid DNA by anion-exchange chromatography in an RNase-free process. Teeters M, Root T, Lightfoot E (1036) Adsorption and desorption behavior of plasmid DNA on ion-exchange membranes: effect of salt valence and compaction agents. doi: 10.1016/j.chroma.2013.03.070Ĭhen WH, Fu JY, Kourentzi K, Willson RC (2011) Nucleic acid affinity of clustered-charge anion exchange adsorbents: effects of ionic strength and ligand density. Mota É, Sousa Â, Černigoj U, Queiroz JA, Tomaz CT, Sousa F (2013) Rapid quantification of supercoiled plasmid deoxyribonucleic acid using a monolithic ion exchanger. Tiainen P, Galaev I, Larsson PO (2007) Plasmid adsorption to anion-exchange matrices: comments on plasmid recovery. Sousa F, Queiroz JA (2011) Supercoiled plasmid quality assessment by analytical arginine-affinity chromatography. Ongkudon CM, Danquah MK (2010) Process optimisation for anion exchange monolithic chromatography of 4.2 kbp plasmid vaccine (pcDNA3F). Matos T, Queiroz JA, Bülow L (2013) Binding and elution behavior of small deoxyribonucleic acid fragments on a strong anion-exchanger multimodal chromatography resin. Yamamoto S, Nakamura M, Tarmann C, Jungbauer A (2007) Retention studies of DNA on anion-exchange monolith chromatography Binding site and elution behavior. Yamamoto S, Yoshimoto N, Tarmann C, Jungbauer A (2009) Binding site and elution behavior of DNA and other large biomolecules in monolithic anion-exchange chromatography. Ghanem A, Healey R, Adly FG (2013) Current trends in separation of plasmid DNA vaccines: a review. Liu MA (2011) DNA vaccines: an historical perspective and view to the future. Pereira LR, Prazeres DMF, Mateus M (2010) Hydrophobic interaction membrane chromatography for plasmid DNA purification: design and optimization. Sousa A, Sousa F, Queiroz JA (2012) Advances in chromatographic supports for pharmaceutical-grade plasmid DNA purification. This behaviour can be linked to the presence of the more hydrophobic phenyl group in Capto Adhere, leading to stronger retention of ssDNA molecules, which have a more hydrophobic character due to a higher degree of base exposure. Another pronounced difference between the resins was observed in the inverted elution of ss- and dsDNA, where ssDNA eluted at 2.88 M NaCl on Capto Adhere, while on Capto Q ImpRes ssDNA eluted already at 1.47 M NaCl. This recognition was not observed for Capto Adhere. Capto Q ImpRes provided a recognition for guanylate bases when samples of deoxynucleotides or poly(dG) were examined. ![]() All deoxynucleotides and DNAs tested bound strongly to the chromatographic materials and could be eluted by a linear gradient of increasing NaCl concentration. These variations in biophysical properties have been utilized for comparative separations on these two resins. The intrinsic differences between single- and double-stranded DNAs are related to charge, hydrophobicity, size and three-dimensional structure. Capto Adhere carries a multimodal ligand which combines strong anion with aromatic recognition, while Capto Q ImpRes is a strong anion exchanger with a chemically similar ligand, but without a phenyl group. Binding of the MAb, purified from CHO cell supernatant by protein A, to Capto adhere ImpRes was therefore performed in 40 mM sodium phosphate, pH 7.8.The differences in chromatographic behaviour of individual deoxynucleotides as well as small single-stranded and double-stranded DNA molecules have been examined for two resins from the Capto family: Capto Adhere and Capto Q ImpRes. The results show that the highest binding capacities were obtained at high pH and low salt concentration, while a shorter residence time reduced the capacity (Figure 1). The influence of pH and NaCl concentration on static binding capacity and dynamic binding capacity of Capto adhere ImpRes was determined in PreDictor™ 96-well filter plates and by design of experiment (DoE) in Tricorn™ 5/50 columns, respectively. Here we describe a workflow for method development of a polishing step for a MAb in B/E mode using Capto adhere ImpRes. The medium allows operation in either bind-elute (B/E) or flow-through (FT) modes. One of these options is Capto adhere ImpRes, a cost-effective and flexible multimodal anion exchange chromatography medium designed for high-resolution polishing of MAbs. GE Healthcare Life Sciences’ MAb toolbox employs protein A chromatography media (resins) as well as a wide range of options for polishing purification steps. A platform approach for MAb purification is desirable as it saves both time and money in process development.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |