AVIPure® AAV Affinity Resins
AVIPure® AAV Affinity Resins provide capture step purification of adeno-associated virus (AAV) 2, 8, and 9 vectors with potential alkaline clean-in-place regeneration for improved process economics. A 50 µm cross-linked agarose matrix bead ensures compatibility with standard bioprocess columns and flow rates.
High performance at a lower price point
AAV process cost reduction
- ~20% lower resin cost/L
- > 20 cycle re-use, NaOH stable
- High binding capacity, yield, purity
Pre-packed or loose resin
- Available now: AAV2, AAV8, AAV9
- Available pre-packed in OPUS® Columns
Secure supply
- Redundant manufacturing at 3+ sites
- Worlds largest ligand manufacturer at 4+ sites
- 10+ year supply agreements
HIGH PERFORMance
High performance at a lower price point
AVIPure® AAV 2, 8, and 9 Affinity Resins meet or exceed the performance metrics of resins developed on alternative platforms with the additional benefit of robust alkaline stability. Affinity resins offer the ability to simplify downstream processing. NaOH stable affinity resins simplify processes and dramatically improve overall process economics.
- Maintain high host cell protein (HCP) clearance and residual DNA clearance levels after 20 cycles
- Clean-in-place (CIP) with 0.5 M NaOH (30 min)
- Retain over 95% of initial yield over 20 use cycles
- >1 x 1014 vp/mL DBC with 1 minute residence time
- >1 x 1015 vp/mL DBC with 4 minute residence time
- High dynamic binding capacity even at short residence time enables use with pre-concentrated feed or direct capture:
- >2 x 1014 vp/mLresin at 1 minute residence time
- >7 x 1014 vp/mLresin at 4 minute residence time
Retain performance with NaOH CIP regeneration
Overlay of 20 bind, wash, elute and CIP cycles using a single column and NaOH based CIP (Concentrated HEK293 CCF lysate). CIP was 30 minutes with 0.5 M NaOH. Exposure time of 0.5 M NaOH exposure time totaled 10 hours.
A bind, wash, elute and CIP cycle was repeated 20 times using concentrated HEK293 clarified cell culture fluid. CIP included a 30-minute hold step with 0.5 M NaOH. Yield averaged 95% across the 20 cycles.
A bind, wash, elute and CIP cycle was repeated 20 times using concentrated HEK293 clarified cell culture fluid. CIP included a 30-minute hold step with 0.5 M NaOH. Residual DNA clearance averaged 5 fold across the 20 cycles.
A bind, wash, elute and CIP cycle was repeated 20 times using concentrated HEK293 clarified cell culture fluid. CIP included a 30-minute hold step with 0.5 M NaOH. Yield averaged 95% and HCP clearance averaged 3.4 logs across the 20 cycles.
Bind with 2-minute residence time, Clear more HCP and DNA
Bind, wash and elute direct capture of AAV cell culture feed (not concentrated) stream using AVIPure® AAV 2 Resin.
A 2 -minute residence time enabled elimination of an existing pre-concentration step. Product was eluted in 50 mM glycine, 150 mM NaCl, 0.01% P188, pH 3 with 83% recovery.
AVIPure® AAV 2 Resin and a competing AAV affinity resin were run under identical conditions. The eluted product was analyzed for purity by SDS-PAGE, HCP and residual DNA.
AVIPure - AAV2 Resin and a competing AAV affinity resin were run under identical conditions (2 minute residence time) with the eluted product analyzed for purity by SDS-PAGE, HCP and residual DNA. AVIPure - AAV2 Resin delivered purified product with 7-fold lower residual HCP, 1.5X lower residual DNA and fewer impurity proteins by SDS-PAGE.
Results
AVIPure® AAV 2 Resin delivered purified product with 7-fold lower residual HCP.
AVIPure® AAV 2 Resin delivered purified product with 1.5X lower residual DNA and fewer impurity proteins by SDS-PAGE.
SPEED TO MARKET
Available pre-packed in OPUS® Columns for quick deployment
AAV affinity resins are available in pre-packed and pre-qualified OPUS® Columns for rapid GMP implementation as well as in loose resin formats.
OPUS® Columns allow you to progress from development to manufacturing scale in weeks, using pre-packed and pre-qualified chromatography columns.
AVIPure® AAV 2, 8, 9 Resin characteristics
|
Category |
Description |
|---|---|
| Base Matrix | Cross-linked agarose, spherical |
| Particle size (d50V) | ~ 50 µm |
| Ligand |
AAV 2 and AAV 8: Alkali-tolerant recombinant protein (animal free) AAV 9: Alkali-tolerant, peptide (synthetic) |
| Coupling chemistry | AAV 2: Thiol AAV 8 and AAV 9: Carbamate |
| Binding capacity |
High dynamic binding capacity even at short residence time enables use with pre-concentrated feed or direct capture: |
| Buffer compatibility | Stable to all commonly used aqueous buffers, including 8 M urea, 6 M guanidine hydrochloride, ethylene glycol, and detergents |
| Solvent compatibility | Water, alcohol (0–20% v/v), acetonitrile, 1–2 M acetic acid, other common organic solvents |
| pH stability | 1-13 |
| Cleaning-in-place stability | 0.1-0.5 M NaOH |
| Pressure/flowa | 3 bar at >300 cm/hr |
| Maximum pressure (ΔP)a | 40 psi |
| Temperature stability | 2–40 °C |
| Delivery conditions | 2% benzyl alcohol |
| Storage | 2–8 °C, 2% benzyl alcohol; do not freeze |
| a In a 2.6 x 20 cm column pressure packed at 4 bar | |
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