Steric exclusion chromatography for purifying large, enveloped viral vectors
Last updated: March 2025
Overview
Lentiviral vectors (LV) are most frequently used as a viral gene delivery platform for the ex vivo generation of chimeric antigen receptor (CAR)-T cells for cancer immunotherapies Steadily increasing demand for LV → development of suitable materials and process strategies for fragile enveloped viral vectors like LV needed. Purification step: one of the major bottlenecks for enveloped viral vectors as elution conditions used in anion exchange chromatography (AEX) and affinity chromatography reduce viral infectivity. Potential alternative for enveloped viral vectors: steric exclusion chromatography (SXC). SXC does not require any chemical interaction between the viral vector and the stationary phase → allows for milder elution conditions and preserves viral activity.
Under optimized conditions, we achieved LV particle recoveries and infectious recoveries of 86% and 88%, respectively. At the same time, high protein and dsDNA removal rates were observed at 81% and 79%, respectively. The developed approach can be used for different viral vector types, regardless of size, shape, and serotype offering a platform approach to purify especially large, fragile enveloped viral vectors.
Download the poster to learn more about the case study.
- Document type: Poster
- Page count: 1
- Read time: 4 minutes
Key Takeaways
- Understanding the feasibility of utilizing steric exclusion chromatography (SXC) for LV purification
- Which process conditions need to be optimized for efficient depletion interaction of the target molecule with the membrane
- How to achieve higher LV particle recoveries and infectious recoveries as well as high protein and dsDNA removal rates
Preview poster
Figure 1. Steric exclusion chromatography as a purification technique for large enveloped viral vectors – lentiviral vectors as a case study.