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Table 4 Reliability of the titration methods to assess lentiviral vector production quality

From: Comparison of lentiviral vector titration methods

packaging plasmid envelope plasmid transfer plasmid RNA/ml TU/ml pg p24/ml
+ + + 6.18 ± 1.71 × 109 1.07 ± 0.53 × 107 4.10 ± 2.05 × 105
+ + - below detection limit below detection limit 2.10 ± 0.64 × 105
+ - + 2.36 ± 0.63 × 1010 below detection limit 1.27 ± 0.32 × 105
- + + 8.04 ± 1.64 × 106 below detection limit below detection limit
  1. Lentiviral vectors were produced in parallel in cell culture dishes by triple transient transfection with transfer, envelope and packaging plasmids. Omission of a plasmid is indicated. RNA equivalents (RNA/ml), transducing units (TU/ml) and p24 concentration (pg p24/ml) were determined after concentration of LV by low-speed centrifugation. For the CH-eGFP-WS vector, the titer was measured with all three methods. In the absence of the packaging plasmid, encoding for structural proteins, the RNA titer decreased 1000-fold while p24 and TU titers were below detection limit. Omission of the envelope plasmid during the vector production resulted in p24 and RNA titers comparable with those of a normal production albeit with a non-detectable functional titer. Vector production without transfer plasmid only yielded a positive p24 titer. Mean values ± standard deviation for 3 measurements of the same sample are shown.