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π-Alpha™ 293 AAV High-yield Production Platform

PackGene’s proprietary π-Alpha™ 293 AAV High-yield Platform uses uniquely designed RC plasmid in the triple-plasmid transfection system to increase AAV production by 3 to 8 times in various AAV serotypes. With combining both in-process upstream and downstream QbD optimizations, the total AAV yield can be increased up to 10 times. The single batch of AAV production delivers up to 1E+17vg virus particles which can meet the needs of most clinical-level and commercial level of AAV production.


  • Increases the AAV yield of AAV in the HEK293 serum-free cell suspension system more than 10 times with uniquely designed RC plasmid and DOE optimizations.
  • The production process increases the suspensive cells production by 10-25 times
  • Unique process development greatly reduces the key impurities (HCD, endotoxin, etc.)
  • Key technology innovations reduce empty capsids rate and increase infection titer

Interested in our AAV services? Please contact us for a free consultation or  submit a request for a quotation.

Intellectual property:

6 patents have been issued with 2 more patents under review

High-yield RC plasmid increases AAV yield by 3-8 times by adding non-coding regulatory elements to the Rep-Cap plasmid (The patent is pending).

Based on DOE experimental optimization of key process parameters, the total AAV yield is increased by more than 10 times

DOE transfection conditions

DOE Engineering Parameters Ambr250 Reactor (Sartorius)

DOE Lysis and harvest conditions

Cell density, total plasmid, plasmid ratio, transfection reagent ratio

Stirring Speed, pH, Dissolved Oxygen, Temperature

Stirring speed, Density, Time, Temperature

AAV production process using serum free 293 suspension cells (200L)


The pilot process development strategy and parameters for the small batches successfully guide the 200L scale-up AAV production process using serum-free 293 suspension cells.

Actual AAV production up to 7.3E+16 vg (total yield) for 200L production system

TEM-Empty capsids<5%

Analytical Ultracentrifuge (AUC)

Lower HCD

Reducing the plasmid DNA residue (Packaged plasmid impurity) significantly by unique molecular modification on plasmid

Reducing the plasmid DNA residue (Packaged plasmid impurity) significantly by unique molecular modification on plasmid

Scalable process by ultracentrifugation allows 200~500L batch production

Characters Descriptions
High yield Design of RC plasmid increases AAV yield by 3-8 times
CPP optimization increases total AAV yields by 10 times
Improved scalability Stable scale-up process: from 125 mL to 200 L (suspension)
Scalable 200 L suspension process: 7.3E+16 vg yeild, >30% recovery rate
Improved biosafety HCD residue: 30~90 ng/1E13 vg (suspension)
Optimal full capsid ratio at harvest (20~66%) and at final (75~94%)
Novel plasmid design decrease 90% encapsidated plasmid impurity

π-Alpha™ 293 AAV High-yield Production Platform