Adenovirus Vector GMP Production Services

Adenovirus vector upstream processes

The adenovirus vector upstream processes comprise 1) mammalian cell culture, e.g. HEK293, A549 in adherent or suspension culture, 2) transduction with adenovirus vector, 3) cell harvest and lysis, and 4) analysis of harvest. The selection of the producer cell line and culture type can be made together with our experts. 

Suspension cell culture – adenovirus vector

Adenovirus vector production in suspension cultures enables easy upscaling and reduces process costs. Producer cell lines such as HEK293 cells can be adapted to suspension culture and they can grow in serum-free media, which enhances the safety profile. For small-scale suspension cultures shake flasks with various capacities can be used. For large-scale suspension cultures, the bioreactor options at Biovian include single-use stirred-tank bioreactors, where the GMP volume is up to 200 L. Depending on the project perfusion, fed-batch or batch cultivation will be applied.

Adherent cell culture

Adenovirus vectors can be produced in adherent cell cultures using T-flasks or multilayer flasks such as hyperflasks or cell stacks. Alternatively, adherent cell cultures can be carried out in bioreactors, where the options at Biovian are packed-bed bioreactors or single-use bioreactors with microcarriers. Adherent cultures require tissue-culture treated vessels and repeated passaging, but allow for easy visual inspection under the microscope.

Adenovirus vector downstream processes – GMP

Biovian’s platform approach to downstream processing of adenovirus vectors comprise 1) tangential flow filtration, TFF, for removal of impurities and buffer exchange 2)  a capture step, where adenovirus vectors are purified and concentrated 3) polishing chromatography 4) concentration and formulation with TFF, or dialysis for low volumes, and 5) analysis of the Drug Substance according to regulatory requirements.

For small-scale production fast gradient ultracentrifugation- based method can be used to purify the clarified adenovirus vector harvest. The final filtration is performed using a 0.2 µm filter to fulfill the sterility requirement for adenovirus DS.

Adenovirus vector Quality Control assays

Biovian offers full access to QC analytics for the demonstration of the quality of adenovirus vector GMP batches. We use compendial assays and perform sample-type specific validation to comply with all regulatory requirements for adenovirus vectors for human use. Development or technical transfer of product-specific analytical methods, as well as validation of these methods, is part of our services.

Adenovirus vector Fill and Finish

Aseptic Fill and Finish of adenovirus vectors is performed in dedicated suites that fulfill BSL 2 requirements. The automated filling line is validated for 2R and 10R vial-sizes up to 1000 vials per session. The tubing, filling needle and other assemblies are fully disposable. In-house release testing services include sterility testing, and a 100% visual inspection is performed after each filling to ensure the safety of the adenovirus vector Drug Product in each vial. Before shipping the adenovirus vector products for use in clinical trials are certified by one of our Qualified Persons. We also provide intermediate GMP warehousing of the Drug Products at various temperatures (RT, +4°C, -20°C and -80°C) on the same European Medicines Agency certified site.

Adenovirus vector GMP documentation package:

• TSE/BSE certificate
• Certificate of analysis
• Batch certificate and certificate of GMP compliance
• GMP production summary report

CMC documentation

• IND/IMPD CMC documentation support upon request

Adenovirus vector stability studies

Stability testing of adenovirus vector Drug Substances and Drug Products is conducted at Biovian under controlled conditions according to ICH guidelines. The stability study services include study planning, analytical QC testing and reporting.