To support the manufacturing of Cell and Gene Therapies, we have developed innovative chromatography solutions, specifically designed to improve the downstream purification of mRNA and viral vectors such as the Adeno-Associated Virus (AAV), lentivirus, and Adv5.
- High selectivity and capacity for the purification of viral vectors and mRNA
- An efficient and robust purification process, from crude material in one step, offering scalability and process consistency
- Significant improvement to the downstream process of viral vectors, by reducing the purification steps and maximizing productivity
- An efficient and scalable solution for the purification of in vitro transcribed mRNA
Our proven CaptureSelect ligand technology, combined with the large pore structure of the POROS beads, enables efficient purification of large biomolecules such as viral vectors. These unique affinity resins are developed to significantly improve downstream processing of viral vectors by reducing the number of purification steps and offering scalability, thus maximizing process consistency, efficiency and productivity.
POROS CaptureSelect AAV resins are unique affinity chromatography resins for the purification of Adeno-associated virus (AAV) used in gene therapy applications.
- High dynamic binding capacity and superior resolution address capacity and selectivity requirements for purifying viral vectors
- Robust viral clearance capabilities of the resins help to reduce the risk of possible viral contamination derived from cell culture production
When the use of your product requires further polishing, such as removal of empty capsids, Thermo Scientific POROS Anion and Cation Exchange resins can be utilized for subsequent polishing steps. The excellent resolution of these resins allow for separation of close related product forms such as empty and full viral capsids.
All AAV affinity purification and polishing resins come with full regulatory support, enabling use in commercial manufacturing.
For more information and products in the AAV manufacturing workflow, please visit our page on AAV production.
Table 1. Summary of POROS CaptureSelect AAV affinity resin properties
|Binding capacity (vg/mL)*||Serotype affinity|
|POROS CaptureSelect AAVX||>1014||AAV1, AAV2, AAV3, AAV4, AAV5. AAV6, AAV7, AAV8, AAV9, recombinant and chimeric vectors**|
|POROS CaptureSelect AAV9||>1014||AAV9|
|POROS CaptureSelect AAV8||>1013||AAV8|
* Measured as viral genomes per milliliter (vg/mL).
** The AAVX Affinity ligand has demonstrated binding reactivity towards a large set of AAV serotypes (including chimeric vectors), suggesting that an even broader range of AAV serotypes might be purified by this resin.
Adenovirus-5 (AdV5) purification
To support the research and development of Adenoviral vectors, a resin for the purification of Adenovirus serotype 5 (AdV5) was developed. POROS CaptureSelect AdV5 Affinity Matrix is specifically designed for the purification and/or scavenging of recombinant hexon capsomeres of AdV5 particles, as well as free hexon proteins. The AdV5 affinity matrix is a research-use-only chromatography resin but can be further developed into a bioprocess resin suitable for cGMP manufacturing.
Lentivirus purification solutions
Heparin affinity chromatography is a simple and effective way of purifying proteins containing a heparin binding domain. Due to its important biological function, the domain is often easy accessible, allowing for efficient purification. POROS 50 HE Heparin Affinity resin can be used for the purification of viral vectors containing a Heparin binding site, such as Lentivirus or Baculovirus.
Alternatively, anion exchange chromatography can serve as an ideal solution for the purification of lentiviral particles. The features of POROS D50 resin, a weak anion exchanger, make it suitable for lentivirus capture.
To support the development of messenger RNA (mRNA)-based vaccines and therapies, a new affinity resin was developed, allowing for efficient and scalable purification of in vitro transcribed (IVT) mRNA. The POROS Oligo (dT) 25 affinity resin was developed to address the selectivity and capacity requirements for large-scale downstream purification of mRNA constructs used in gene therapy applications. This purification solution effectively separates polyadenylated (polyA) mRNA from components of the transcription reaction process using simple salt and water purification steps. The POROS Oligo (dT) 25 affinity resin helps to simplify the mRNA purification workflow by reducing the complexity of subsequent polish steps.
Additional reading and webinars
- Accelerating Advancement in Gene Therapy by Improving Downstream Purification of Viral Vectors
- AAV vector process development achieving high purity & high yield – experiences from the front line
- AAV downstream processing for industrial scale production - moving towards commercial manufacturing
- Combining state-of-the-art production, purification, and analytics to optimize AAV manufacturing for clinical and commercial gene therapies