11
3.3 DNA Delivery Into Cells Using
Electroporation (cont’d)
3.3.2 AgilePulse
™
Electroporation
In section 3.3.1 general concepts of electroporation were
presented. This section presents the types of waveforms and
electrodes available for use in the AgilePulse
™
system.
Waveforms
Traditional in vivo electroporation uses four to six rectangular
pulses that are 100 microseconds in duration at a rate of one
per second. Thus the total treatment time is four to six seconds.
More advanced waveforms such as the PulseAgile
®
have been
found to be more effective in eliciting T-cell and antibody
responses simultaneously when delivering DNA vaccines (Roos
et al, 2006, Vertuani et al 2009, Bråve et al 2009). A PulseAgile
®
waveform consists of various pulse groups with different
characteristics from group to group. For example, the study by
Dr. Roos found the optimum waveform for induction of high
gene expression and high induction of antigen-specific T cells to
be:
The total treatment time for this waveform is 0.27 seconds (this
waveform is currently tested in three clinical studies). When the
pulses are delivered in less than half a second there is only one
muscle contraction and tolerability is highly improved.
The AgilePulse
™
system has the following waveform settings
available:
Three groups may be used. A group is 1-10 pulses with the same
parameters. The system software automatically concatenates
each group.
Group Pulse
Amplitude
Pulse
Width
Pulse
Interval
Group
Interval
Pulse
Number
1
450 V
(1125 V/cm)
0.05 ms 300 ms 500 ms 2
2
110 V
(275 V/cm)
10 ms
300 ms 500 ms 8
Pulse
Amplitude
Pulse
Width
Pulse
Interval*
Maximum
Duty Cycle
Number
of Pulses
50 to
300 V
0.050 to
10 ms
0.2 to
1000 ms
50%
10
310 to 1000
V
0.050 to
1 ms
0.2 to
1000 ms
50%
10
Group Pulse
Amplitude
Pulse
Width
Pulse
Interval
Group
Interval
Pulse
Number
1
450 V
(1125 V/cm)
0.05 ms 0.2 ms 50 ms
2
2
110 V
(275 V/cm)
10 ms
20 ms
50 ms
8
*Limited to 50% Duty Cycle, i.e., (Pulse Width/Pulse Interval) less than or
equal to 0.5
The total treatment time for this waveform is 2.98 seconds.
(Waveforms for clinical use must be optimized in pre-clinical
studies.)
Another more recent study by Roos et al (Mol. Ther., 2009)
developed a variant of the PulseAgile
®
waveform described
above. This waveform is called Fast PulseAgile
®
(PAfast) and
differs only in the length of the pulse intervals:
AgilePulse
™
Tutorial
3
