Johnson, S;
Wheeler, JX;
Thorpe, R;
Collins, M;
Takeuchi, Y;
Zhao, Y;
(2018)
Mass spectrometry analysis reveals differences in the host cell protein species found in pseudotyped lentiviral vectors.
Biologicals
, 52
pp. 59-66.
10.1016/j.biologicals.2017.12.005.
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Abstract
Lentiviral vectors (LVs) have been successfully used in clinical trials showing long term therapeutic benefits. Studying the role of cellular proteins in lentivirus HIV-1 life cycle can help understand virus assembly and budding, leading to improvement of LV production for gene therapy. Lentiviral vectors were purified using size exclusion chromatography (SEC). The cellular protein composition of LVs produced by two different methods was compared: the transient transfection system pseudotyped with the VSV-G envelope, currently used in clinical trials, and a stable producer cell system using a non-toxic envelope derived from cat endogenous retrovirus RD114, RDpro. Proteins of LVs purified by size exclusion chromatography were identified by tandem mass spectrometry (MS/MS). A smaller number of cellular protein species were detected in stably produced vectors compared to transiently produced vector samples. This may be due to the presence of co-purified VSV-G vesicles in transiently produced vectors. AHNAK (Desmoyokin) was unique to RDpro-Env vectors. The potential role in LV particle production of selected proteins identified by MS analysis including AHNAK was assessed using shRNA gene knockdown technique. Down-regulation of the selected host proteins AHNAK, ALIX, and TSG101 in vector producer cells did not result in a significant difference in vector production.
| Type: | Article |
|---|---|
| Title: | Mass spectrometry analysis reveals differences in the host cell protein species found in pseudotyped lentiviral vectors |
| Location: | England |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1016/j.biologicals.2017.12.005 |
| Publisher version: | https://doi.org/10.1016/j.biologicals.2017.12.005 |
| Language: | English |
| Additional information: | Copyright © 2017 Published by Elsevier Ltd on behalf of International Alliance for Biological Standardization. Available under a Creative Commons Attribution-NonCommercial-NoDerivatives (CC BY-NC-ND 4.0) licence (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
| Keywords: | AHNAK, ALIX, Host proteins, Lentiviral vectors, Mass spectrometry |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Infection and Immunity |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10042724 |
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