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Title: Comparison of lentiviral vector titration methods
Authors: Geraerts, Martine
Willems, Sofie
Baekelandt, Veerle
Debyser, Zeger ×
Gijsbers, Rik #
Issue Date: Jul-2006
Publisher: BioMed Central
Series Title: BMC Biotechnology vol:6
Article number: 34
Abstract: BACKGROUND: 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. RESULTS: 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. CONCLUSION: The different titration methods have specific advantages and disadvantages. Depending on the experimental set-up one titration method should be preferred over the others.
URI: 
ISSN: 1472-6750
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Molecular Virology and Gene Therapy
Interdepartemental Stem Cell Institute (-)
Research Group for Neurobiology and Gene Therapy
× corresponding author
# (joint) last author

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