Comparison of lentiviral vector titration methods
<p>Abstract</p> <p>Background</p> <p>Lentiviral vectors are efficient vehicles for stable gene transfer in dividing and non-dividing cells. Several improvements in vector design to increase biosafety and transgene expression, have led to the approval of these vectors fo...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2006-07-01
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| Series: | BMC Biotechnology |
| Online Access: | http://www.biomedcentral.com/1472-6750/6/34 |
| _version_ | 1818156007610646528 |
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| author | Debyser Zeger Baekelandt Veerle Willems Sofie Geraerts Martine Gijsbers Rik |
| author_facet | Debyser Zeger Baekelandt Veerle Willems Sofie Geraerts Martine Gijsbers Rik |
| author_sort | Debyser Zeger |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Lentiviral vectors are efficient vehicles for stable gene transfer in dividing and non-dividing cells. Several improvements in vector design to increase biosafety and transgene expression, have led to the approval of these vectors for use in clinical studies. Methods are required to analyze the quality of lentiviral vector production, the efficiency of gene transfer and the extent of therapeutic gene expression.</p> <p>Results</p> <p>We compared lentiviral vector titration methods that measure pg p24/ml, RNA equivalents/ml, transducing units (TU/ml) or mRNA equivalents. The amount of genomic RNA in vector particles proves to be reliable to assess the production quality of vectors encoding non-fluorescent proteins. However, the RNA and p24 titers of concentrated vectors are rather poor in predicting transduction efficiency, due to the high variability of vector production based on transient transfection. Moreover, we demonstrate that transgenic mRNA levels correlate well with TU and can be used for functional titration of non-fluorescent transgenes.</p> <p>Conclusion</p> <p>The different titration methods have specific advantages and disadvantages. Depending on the experimental set-up one titration method should be preferred over the others.</p> |
| first_indexed | 2024-12-11T14:51:27Z |
| format | Article |
| id | doaj.art-e84f778196274862a5a5e997451e1991 |
| institution | Directory Open Access Journal |
| issn | 1472-6750 |
| language | English |
| last_indexed | 2024-12-11T14:51:27Z |
| publishDate | 2006-07-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Biotechnology |
| spelling | doaj.art-e84f778196274862a5a5e997451e19912022-12-22T01:01:28ZengBMCBMC Biotechnology1472-67502006-07-01613410.1186/1472-6750-6-34Comparison of lentiviral vector titration methodsDebyser ZegerBaekelandt VeerleWillems SofieGeraerts MartineGijsbers Rik<p>Abstract</p> <p>Background</p> <p>Lentiviral vectors are efficient vehicles for stable gene transfer in dividing and non-dividing cells. Several improvements in vector design to increase biosafety and transgene expression, have led to the approval of these vectors for use in clinical studies. Methods are required to analyze the quality of lentiviral vector production, the efficiency of gene transfer and the extent of therapeutic gene expression.</p> <p>Results</p> <p>We compared lentiviral vector titration methods that measure pg p24/ml, RNA equivalents/ml, transducing units (TU/ml) or mRNA equivalents. The amount of genomic RNA in vector particles proves to be reliable to assess the production quality of vectors encoding non-fluorescent proteins. However, the RNA and p24 titers of concentrated vectors are rather poor in predicting transduction efficiency, due to the high variability of vector production based on transient transfection. Moreover, we demonstrate that transgenic mRNA levels correlate well with TU and can be used for functional titration of non-fluorescent transgenes.</p> <p>Conclusion</p> <p>The different titration methods have specific advantages and disadvantages. Depending on the experimental set-up one titration method should be preferred over the others.</p>http://www.biomedcentral.com/1472-6750/6/34 |
| spellingShingle | Debyser Zeger Baekelandt Veerle Willems Sofie Geraerts Martine Gijsbers Rik Comparison of lentiviral vector titration methods BMC Biotechnology |
| title | Comparison of lentiviral vector titration methods |
| title_full | Comparison of lentiviral vector titration methods |
| title_fullStr | Comparison of lentiviral vector titration methods |
| title_full_unstemmed | Comparison of lentiviral vector titration methods |
| title_short | Comparison of lentiviral vector titration methods |
| title_sort | comparison of lentiviral vector titration methods |
| url | http://www.biomedcentral.com/1472-6750/6/34 |
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