ArticleGene therapy2001
A highly efficient and consistent method for harvesting large volumes of high-titre lentiviral vectors.
Article in Gene therapy, 2001. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 65 citations in OpenAlex.
- Lentiviral Vector Bioprocessing.Viruses · 2021Review
- Lentivirus Mediated Pancreatic Beta-Cell-Specific Insulin Gene Therapy for STZ-Induced Diabetes.Molecular therapy : the journal of the American Society of Gene Therapy · 2021Article
- Therapeutic Effects of (5R)-5-Hydroxytriptolide on Fibroblast-Like Synoviocytes in Rheumatoid ArthritisFrontiers in immunology · 2021Article
- A new chemical complex can rapidly concentrate lentivirus and significantly enhance gene transduction.Cytotechnology · 2018Article
- An HNF1α-regulated feedback circuit modulates hepatic fibrogenesis via the crosstalk between hepatocytes and hepatic stellate cells.Cell research · 2015Article
- MGMT enrichment and second gene co-expression in hematopoietic progenitor cells using separate or dual-gene lentiviral vectors.Virus research · 2015Article
- Production, purification and titration of a lentivirus-based vector for gene delivery purposes.Cytotechnology · 2014Article
- Promoter Targeting shRNA Suppresses HIV-1 Infection In vivo Through Transcriptional Gene Silencing.Molecular therapy. Nucleic acids · 2013Article
- Simplified production and concentration of lentiviral vectors to achieve high transduction in primary human T cells.BMC biotechnology · 2013Article
- Estradiol protects dermal hyaluronan/versican matrix during photoaging by release of epidermal growth factor from keratinocytes.The Journal of biological chemistry · 2012Article
- A bidirectional promoter architecture enhances lentiviral transgenesis in embryonic and extraembryonic stem cells.Gene therapy · 2011Article
- Inhibition of oesophageal squamous cell carcinoma progression by in vivo targeting of hyaluronan synthesis.Molecular cancer · 2011Article
- Neonatal gene therapy of glycogen storage disease type Ia using a feline immunodeficiency virus-based vector.Molecular therapy : the journal of the American Society of Gene Therapy · 2010Article
- Lentiviral-mediated RNA interference against TGF-beta receptor type II in renal epithelial and fibroblast cell populations in vitro demonstrates regulated renal fibrogenesis that is more efficient than a nonlentiviral vector.Journal of biomedicine & biotechnology · 2010Article
- Dendritic cells for active anti-cancer immunotherapy: targeting activation pathways through genetic modification.Endocrine, metabolic & immune disorders drug targets · 2009Review
- Streamlined design of a self-inactivating feline immunodeficiency virus vector for transducing ex vivo dendritic cells and T lymphocytes.Genetic vaccines and therapy · 2007Article
- Lentiviral vectors for cancer immunotherapy: transforming infectious particles into therapeutics.Gene therapy · 2007Review
- Conjugation of lentivirus to paramagnetic particles via nonviral proteins allows efficient concentration and infection of primary acute myeloid leukemia cells.Journal of virology · 2005Article
- Downstream processing of viral vectors and vaccines.Gene therapy · 2005Review
- Altering the tropism of lentiviral vectors through pseudotyping.Current gene therapy · 2005Review
Corrections and comments
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Authors and funding
6 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Lentiviral vectors pseudotyped with vesicular stomatitis virus glycoprotein (VSV-G) are emerging as the vectors of choice for in vitro and in vivo gene therapy studies. However, the current method for harvesting lentivectors relies upon ultracentrifugation at 50,000 g for 2 h. At this ultra-high speed, rotors currently in use generally have small volume capacity. Therefore, preparations of large volumes of high-titre vectors are time-consuming and laborious to perform. In the present study, viral vector supernatant harvests from vector-producing cells (VPCs) were pre-treated with various amounts of poly-L-lysine (PLL) and concentrated by low speed centrifugation. Optimal conditions were established when 0.005% of PLL (w/v) was added to vector supernatant harvests, followed by incubation for 30 min and centrifugation at 10,000 g for 2 h at 4 degrees C. Direct comparison with ultracentrifugation demonstrated that the new method consistently produced larger volumes (6 ml) of high-titre viral vector at 1 x 10(8) transduction unit (TU)/ml (from about 3,000 ml of supernatant) in one round of concentration. Electron microscopic analysis showed that PLL/viral vector formed complexes, which probably facilitated easy precipitation at low-speed concentration (10,000 g), a speed which does not usually precipitate viral particles efficiently. Transfection of several cell lines in vitro and transduction in vivo in the liver with the lentivector/PLL complexes demonstrated efficient gene transfer without any significant signs of toxicity. These results suggest that the new method provides a convenient means for harvesting large volumes of high-titre lentivectors, facilitate gene therapy experiments in large animal or human gene therapy trials, in which large amounts of lentiviral vectors are a prerequisite.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.