AVB Sepharose High Performance

• Cat#No#: AV-341C
• Product Overview: AVB Sepharose High Performance is an affinity resin designed for the purification of adeno associated virus (AAV), Fast, one-step purification of adeno associated viruses of several subclasses:
  One step purification of adeno associated viruses.
  Affinity resin with high selectivity and low background binding.
  Large surface area to give enhanced capacity.
  Prepacked HiTrap columns available, for simple operation with a syringe, pump, or chromatographic system.
• Description: AVB Sepharose High performance is an affinity resin with affinity for adeno associated viruses. It enables purification of adeno associated viruses in one step with high purity and excellent yields. It has proven affinity for adeno associated viruses from subclasses 1, 2, 3, and 5, but it is expected to have affinity for most of the other adeno associated virus subclasses. The ligand is a 14 kD fragment from a single chain antibody, expressed in yeast.
• Characteristic: Efficient, industrial-scale purification of adeno-associated viruses (AAV) of several subclasses by affinity chromatography.
  Reduced regulatory concerns (due to non-mammalian derived product) in the production of AAV for clinical applications.
  High selectivity and excellent scalability.
• Applications: When using AVB Sepharose High Performance the AAV can be applied directly from clarified AAV vector cell lysate. Conventional buffers (e.g., PBS, Tris, citrate) may be used for loading, washing, and elution. Virus binds to the column at around neutral pH and is typically eluted by lowering the pH, for example in the range of pH 2 to 5. Since AAV is sensitive to highly acidic conditions, it is important to minimize the exposure to low pH during elution. Therefore, collected elution fractions should be neutralized immediately.
• Maximum operating pressure: 100-200 cm/h, 300 kPa, BioPilot 60/600 column, bed height 30 cm.
• Ligand Coupling Method: Amide linkage
• Matrix: Cross-linked agarose
• Particle Size: 24 µm-44 µm
• Average particle size: ~34 µm
• Ligand: 14 kD recombinant protein
• Ligand density: Approx. 2.4 mg/ml of medium
• Dynamic binding capacity: > 10^12 genecopies / mL
• Recommended flow rate: 100-200 cm/h, 300 kPa, BioPilot 60/600 column, bed height 30 cm.
• Recommended column height: 30 cm
• Chemical stability: Stable to commonly used aqueous buffers.
• Physical stability: The ligand is linked to the Sepharose High Performance base matrix via a stable amide bond.
• Storage: 4 to 8°C, 20% Ethanol
• Cleaning-in-place: The recommended protocol comprises initial strip of the resin at low pH, and then subjecting the resin to NaOH of low concentration for cleaning.
• Sanitization: PAB (120 mM phosphoric acid, 167 mM acetic acid, 2.2% v/v benzyl alcohol) is used for final sanitization of the resin.
• Pack size: 75 mL
• Maximum flow velocity: 200 cm/h
• Wash buffer: 20 mM Tris-HCl, 0.5 M NaCl, pH 8.0