Implementation of Novel Affinity Ligand for Lentiviral Vector Purification

Ana Sofia Moreira, Sandra Bezemer, Tiago Q. Faria, Frank Detmers, Pim Hermans, Laurens Sierkstra, Ana Sofia Coroadinha, Cristina Peixoto

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)
29 Downloads (Pure)

Abstract

The use of viral vectors as therapeutic products for multiple applications such as vaccines, cancer treatment, or gene therapies, has been growing exponentially. Therefore, improved manufacturing processes are needed to cope with the high number of functional particles required for clinical trials and, eventually, commercialization. Affinity chromatography (AC) can be used to simplify purification processes and generate clinical-grade products with high titer and purity. However, one of the major challenges in the purification of Lentiviral vectors (LVs) using AC is to combine a highly specific ligand with a gentle elution condition assuring the preservation of vector biological activity. In this work, we report for the first time the implementation of an AC resin to specifically purify VSV-G pseudotyped LVs. After ligand screening, different critical process parameters were assessed and optimized. A dynamic capacity of 1 × 1011 total particles per mL of resin was determined and an average recovery yield of 45% was found for the small-scale purification process. The established AC robustness was confirmed by the performance of an intermediate scale providing an infectious particles yield of 54%, which demonstrates the scalability and reproducibility of the AC matrix. Overall, this work contributes to increasing downstream process efficiency by delivering a purification technology that enables high purity, scalability, and process intensification in a single step, contributing to time-to-market reduction.

Original languageEnglish
Article number3354
JournalInternational Journal of Molecular Sciences
Volume24
Issue number4
DOIs
Publication statusPublished - Feb 2023

Keywords

  • affinity chromatography
  • cell therapy
  • gene therapy
  • lentiviral vector
  • viral vector manufacturing

Fingerprint

Dive into the research topics of 'Implementation of Novel Affinity Ligand for Lentiviral Vector Purification'. Together they form a unique fingerprint.

Cite this