UCL Discovery
UCL home » Library Services » Electronic resources » UCL Discovery

Time-dependent sorption behavior of lentiviral vectors during anion-exchange chromatography

Pamenter, George; Davies, Lee; Knevelman, Carol; Miskin, James; Mitrophanous, Kyriacos; Dikicioglu, Duygu; Bracewell, Daniel G; (2023) Time-dependent sorption behavior of lentiviral vectors during anion-exchange chromatography. Biotechnology and Bioengineering 10.1002/bit.28483. (In press). Green open access

[thumbnail of Time dependent sorption behavior of lentiviral vectors during anion exchange.pdf]
Preview
Text
Time dependent sorption behavior of lentiviral vectors during anion exchange.pdf - Published Version

Download (3MB) | Preview

Abstract

Use of lentiviral vectors (LVs) in clinical Cell and Gene Therapy applications is growing. However, functional product loss during capture chromatography, typically anion-exchange (AIEX), remains a significant unresolved challenge for the design of economic processes. Despite AIEX's extensive use, variable performance and generally low recovery is reported. This poor understanding of product loss mechanisms highlights a significant gap in our knowledge of LV adsorption and other types of vector delivery systems. This work demonstrates HIV-1-LV recovery over quaternary-amine membrane adsorbents is a function of time in the adsorbed state. Kinetic data for product loss in the column bound state was generated. Fitting a second order-like rate model, we observed a rapid drop in functional recovery due to increased irreversible binding for vectors encoding two separate transgenes ( t Y 1 / 2 ${t}_{{Y}_{1/2}}$  = 12.7 and 18.7 min). Upon gradient elution, a two-peak elution profile implicating the presence of two distinct binding subpopulations is observed. Characterizing the loss kinetics of these two subpopulations showed a higher rate of vector loss in the weaker binding peak. This work highlights time spent in the adsorbed state as a critical factor impacting LV product loss and the need for consideration in LV AIEX process development workflows.

Type: Article
Title: Time-dependent sorption behavior of lentiviral vectors during anion-exchange chromatography
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.1002/bit.28483
Publisher version: https://doi.org/10.1002/bit.28483
Language: English
Additional information: © 2023 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Keywords: Cell and Gene Therapy, capture chromatography, downstream processing, lentiviral purification, lentivirus, viral vector
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/10172823
Downloads since deposit
35Downloads
Download activity - last month
Download activity - last 12 months
Downloads by country - last 12 months

Archive Staff Only

View Item View Item