Skip to Content
Merck
CN

Viral Vector Downstream Processing

Viral vector purification processes have historically been based on systems used for monoclonal antibodies, which are not designed to deliver the necessary level of recovery.

The introduction of new technologies specifically designed for gene therapy viral vector downstream processing offers the opportunity to increase yield and throughput. Among the approaches that can be leveraged to optimize viral vector downstream processing include:

  • Ultrafiltration/diafiltration operations, particularly with the use of tangential flow filtration (TFF) for cell or viral harvesting and purification, downstream protein concentration and diafiltration, as well as in final formulation
  • Intensified chromatography processes that reduce the number and size of unit operations, via technologies such as high-productivity, single-use chromatography membrane adsorbers

These downstream technologies offer the potential to reduce viral vector purification times from hours to minutes, while improving recovery. In addition, they can facilitate scale-up, reduced the process footprint, and enable more efficient facility utilization.


Related Technical Articles

Find More Articles


Primary Clarification

The crude medium containing viral vectors is depth filtration or a combination of both centrifugation and depth filtration to remove large cellular debris and other contaminants.

Secondary Clarification

Clarified medium from the primary clarification is further purified through additional steps.

Ultrafiltration/Diafiltration

Ultrafiltration and diafiltration are used for concentration and buffer exchange, respectively. The viral vector is separated from impurities using size-based filtration. Diafiltration removes salt and other molecules, replacing with the buffer of choice.

Chromatography

At least two chromatography methods are used to purify the viral vector including ion exchange, affinity and size exclusion.

Virus Filtration

The purified viral vector is subjected to nanofiltration with 15 nanometers pores to remove adventitious viruses, endotoxins, and any other remaining impurities and aggregates.

Sterile Filtration

Sterile filtration, or bioburden reduction, can be used multiple times in the process. The feed stream is passed through a 0.4 or 0.2 micron sterilizing grade filter to remove bacteria and other microorganisms that have been introduced during the processing.

Final Fill

The purified and sterile viral vector is filled into the final intended container. Careful considerations need to be given here for formulation, packaging materials, storage condition to ensure stability and integrity.


Related Resources


Workflow

Colorful illustration of four simplified characters against a yellow background. Left to right: doctor with large pink model, child, and two adults.

Gene Therapy Manufacturing

The rise of gene therapies is driving rapid innovation, but manufacturers face complex challenges bringing new therapies to life

Simplified line drawing of equipment used in processing

Making the right upstream process decisions not only impacts viral vector titer, but downstream processes, timelines, and regulatory acceptance

A purple outline of a chemical flask connected to a condenser, depicted against a white background.

Formulating a commercially viable gene therapy demands a high level of application and regulatory expertise

A purple line drawing of a magnifying glass focusing on a pattern symbolizing molecular structure..

Critical biosafety testing and characterization of viral vector products can help to fully analyze key quality attributes: identity, potency, safety, and stability

A purple line drawing of three bottles on a conveyor belt, with the middle bottle being filled by a nozzle.

CDMO partnerships play a critical role in advancing clinical pipelines and achieving successful commercialization




Sign In To Continue

To continue reading please sign in or create an account.

Don't Have An Account?