AVIPure® - AAV2, 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 can both simplify processes and dramatically improve overall process economics. All three AAV affinity resins have been shown to retain over 95% of their dynamic binding capacity (DBC), host cell protein (HCP) clearance and residual DNA clearance after 20 cycles with a 0.5 M NaOH (30 min) clean-in-place (CIP) step.
Bind, wash, and, elute, trace for direct capture of AAV cell culture feed (not concentrated) stream using AVIPure® - AAV2 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® - 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.
|AVIPure® - AAV2 Resin||4400 ppm||2400 ppm|
|Supplier A||33000 ppm||4000 ppm|
|2||AVIPure® NaOH CIP|
|4||Competitor NaOH CIP|
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.
|Base Matrix||Cross-linked agarose, spherical|
|Particle size (d50V)||~ 50 µm|
AAV2 and AAV8: Alkali-tolerant recombinant protein (animal free)
AAV9: Alkali-tolerant, peptide (synthetic)
|Coupling chemistry||AAV2: Thiol
AAV8 and AAV9: Carbamate
Achieve high DBC
>1 x 1014 vp/mL DBC with 1 minute residence time
>1 x 1015 vp/mL DBC with 4 minute residence time
|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|
|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|
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.
ABOUT OPUS® COLUMNS
Repligen develops and manufactures products for the biopharmaceutical industry. We focus on the timely delivery of high quality, consistent and robust products, to ensure business continuity for our customers.
Avitide, a Repligen company, is an ISO 9001:2015 certified resin manufacturer and supplier of custom affinity purification resins based in Lebanon, NH. Redundancy is built into the manufacturing model across the U.S. and Europe, with multiple resin warehousing locations, multiple qualified resin production facilities, and multiple ligand production facilities.