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Description
POROS CaptureSelect AAV9 high performance affinity resin is engineered to address the high selectivity and capacity requirements for the purification of viral vectors used for gene therapy applications and is specifically intended for the purification of adeno-associated virus type 9 (AAV9).
Features of POROS CaptureSelect AAV9 resin include:
- One-step AAV9 purification from crude material with high purity and yield
- High specificity and capacity, which helps reduce the process volume significantly for subsequent steps and maximizes yield
- Low operating back pressure and linear pressure versus flow responses driving flexible scalability
- Superior capacity maintained at high flow rates
- Rigid polymeric bead with covalent surface chemistry for easier handling and packing, with superior physical and chemical stability enabling a more robust downstream purification process
- Non-animal derived—the ligand is a 13-kDa single-domain fragment that comprises the 3 CDRs that form the antigen-binding domain and is efficiently produced by the yeast Saccharomyces cerevisiae in a production process free of animal components.
POROS CaptureSelect AAV9 affinity resin specifically binds AAV9 and enables a robust, fast, and efficient purification process with excellent purity and yield obtained in one step. The use of this affinity resin will enable significant improvement to downstream processing by reducing the purification steps and maximizing productivity, for scalability and processing consistency.
Main characteristics:
- Matrix: cross-linked poly(styrene-divinylbenzene)
- Average particle size: 50 μm
- Ligand: POROS CaptureSelect AAV9 affinity ligand
- Ligand coupling method: carbonyldiimidazole (CDI)
- Binding capacity: >1 x 1014 genome copies/mL of resin
- Elution conditions: acidic elution with 0.1 M glycine, pH 3.0
- Mechanical resistance: 100 bar (1450 psi; 10 MPa)
- Formulation buffer: 18% (v/v) ethanol
Specifications
Specifications
| Binding Capacity | >1 x 1014 genome copies/mL of resin |
| Matrix | Cross-linked poly(styrene-divinylbenzene) |
| Content And Storage | Store at 2°C to 8°C. |
| Shipping Condition | Wet Ice |
| Particle Size | 50 μm |
| Form | Liquid Suspension |
| Product Line | CaptureSelect, POROS |
| Quantity | 10 mL |
| Type | Resin |
| Ligand Type | AAV9 affinity ligand |
Frequently Asked Questions (FAQs)
All of the pre-packed POROS columns, regardless of column size or resin, have the same 10-32 female fittings on the column. Columns are compatible with Waters and Agilent HPLC systems. You will need 1/16 inch male fittings for the HPLC to attach the column.
All of the pre-packed columns, regardless column size or resin, have the same 10-32 female fittings on the column. They are all compatible with the AKTA instrument platform, however due to the column volume of the 2.1 mmD x 30 mmL columns (0.1 mL), these will not perform well in AKTA or Avant purification systems due to the relative size of the hold- up volume in these systems. The smallest column (PEEK or SS) that could feasibly be used in these systems is the 4.6mmD x 50mmL (0.8 mL) format.
The physical stability of POROS resins is high. Customers have agitated, with overhead mixing and or bottom gassing for long periods of time (5-10 hours) and have performed hundreds of diaphragm pump passes during stall packs. Keep in mind that agitation with any device that creates grinding and sheer, such as with a magnetic stir bar, is not recommended for any type of resin.
POROS resins rehydrate quite well. Here is the procedure that we recommend:
1. Flow 5% ethanol over the column at ?350 cm/hour for 3 CVs, followed by water or 0.1 M sodium chloride at 300 cm/hour for 3 CVs.
2. If baseline noise is still present, flow 20% ethanol over the column at 300 cm/hour for 3 CVs, followed by water or 0.1 M sodium chloride at 300 cm/hour for 3 CVs.
3. If you have established column qualification tests and specifications, perform the tests after rewetting.
In general POROS resins have behaved better for large biomolecules such as viral particles, fusion proteins, and globular proteins, where shape (not size) matters due to its unique pore structure and large pore structure as compared to traditional resins. As the target molecule size increases, capacities obtained will decrease. The large pore structure of POROS resins, which allows for convective flow (and therefore enhanced diffusion) is especially well suited for the purification of large biomolecules. POROS resins offer the best blend of high capacity (associated with chromatography beads) and improved chromatography efficiency (typically associated with monoliths or membranes). The average pore size for POROS resins is 1,000-3,600 Angstrom (100-360 nm) depending on the base bead/chemistry being utilized. For example, the molecular weight of IgG is 150 kDa and the molecular radius is 55 Angstrom. The molecular weight of IgM is ~900 kDa (pentamer), and the molecular radius is 120 Angstrom. So both of these biomolecules can interact with the pore structure associated with POROS materials.
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