PlasmidSelect Xtra

• Cat#No#: Pl-427C
• Product Overview: PlasmidSelect Xtra is a thiophilic aromatic adsorption chromatography resin with a selectivity that allows supercoiled covalently closed circular forms of plasmid DNA to be separated from open circular forms.
  PlasmidSelect Xtra allows for generic purification of supercoiled plasmid DNA.
  PlasmidSelect Xtra allows for consistent purification results when scaling up from research scale to cGMP production.
  Available in convenient prepacked formats for research, process development and quantitative and qualitative analysis of supercoiled plasmid DNA.
  The hydrophilic nature of the base matrix ensures low levels of non-specific binding.
  This resin is a BioProcess resin supported for industrial applications.
• Description: PlasmidSelect Xtra chromatography medium forms the basis of a generic process for purifying supercoiled (sc) covalently closed circular plasmid DNA suitable for bulk to clinical-grade applications. The process provides high capacity, delivers high yields, and can be scaled up to fulfill requirements for the economical industrial manufacture of plasmid DNA in highly regulated environments. The same principle can also be used to rapidly analyze the quantity and quality of plasmid DNA in complex solutions.
• Characteristic: Generic process for purification of supercoiled plasmid DNA.
  Consistent from research to cGMP manufacturing.
  Screening kit: Quick and easy analysis with an ÄKTA chromatography system.
  Starter kit: Prepacked columns for convenient process development.
  Bulk medium: PlasmidSelect Xtra medium is a BioProcess medium available in large quantities for scale-up and manufacture.
• Maximum operating pressure: < 120 cm/h, XK 16/20 column.
• Sample preparation: The starting material for plasmid DNA (pDNA) is usually clarified bacterial cell lysate containing the desired plasmid1. Generally, fresh or frozen cell paste is suspended in 50 mM Tris-HCl buffer, pH 7.5, containing 10 mM EDTA to reduce DNAse activity and approximately 50 mM glucose. Lyse cells by adding 0.2 M NaOH, 1% SDS at room temperature. The suspension normally changes color and becomes viscous.
  Flocculate cellular debris and SDS complexes by gently adding cold 3 M potassium acetate, pH 5 and incubating on ice. Clarify the plasmid DNA extract using filtration or centrifugation. This procedure normally gives an initial concentration of approximately 0.05 to 0.1 mg/ml plasmid DNA in an alkaline lysate.
• Matrix: cross-linked agarose
• Particle Size: 24 µm-44 µm
• Average particle size: ~34 µm
• Ligand: 2-mercaptopyridine
• Ligand density: Approx. 3.5 mg 2-mercaptopyridine/ml medium.
• Dynamic binding capacity: ≤ 120 cm/h
• Recommended flow rate: 30 to 300 cm/h (2.7 to 27 ml/min)
• Recommended column height: 100 mm
• Chemical stability: Stable in commnly used aqueous buffers
• pH working range: 3–11
• CIP stability: 2–13
• Temperature stability: 15°C to 30°C
• Storage: 4 to 30°C, 20% Ethanol
• Cleaning-in-place: 1. Wash with at least 10 to 15 ml (2 to 3 CV) of water at a flow rate of 4 ml/min (120 cm/h).
  2. Wash with 10 to 15 ml (2 to 3 CV) of 0.5 M NaOH at a low flow rate of 1.3 ml/min (40 cm/h).
  3. Incubate the column for at least 15 min.
  4. Wash with at least 10 to 15 ml (2 to 3 CV) of water at a flow rate of 4 ml/min (120 cm/h).
  5. Re-equilibrate the column with 10 to 15 ml (2 to 3 CV) of Buffer B.
• Pack size: 25 mL
• BioProcess resin: Yes
• Maximum flow velocity: 450 cm/h (40 ml/min)
• Dimensions: 26 mm
• Column volume: 53 ml
• Column hardware pressure limit: 0.5 MPa, 5 bar, 72 psi