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Hybrimune Hybridoma Production System
Publication 015-1010191 Rev 4.0 • www.btxonline.com
Introduction
The Hybrimune™ Hybridoma Production System was developed
for large volume commercial applications using cell electrofusion.
This includes hybridoma production and tumor/dendritic cell
immunotherapy. The system has not been approved for clinical or
in vitro diagnostic use. The system can fuse up to 180 million cells
in one process run. The Hybrimune is designed to be simple to set-
up and to operate. The system is shown in Figure 1.
The Hybrimune System is comprised of four components: the
Hybrimune Waveform Generator, the user-interface software,
two large volume coaxial chambers (2 ml optimization chamber
and 9 ml production chamber), and the low conductivity BTXpress
Cytofusion® Medium C in which the electrofusion takes place.
These components were all designed to operate as a system. Using
other components may damage the system and may result in
degraded performance. A computer is required for the application
software but is not included in the system.
The Hybrimune Hybridoma Production System is the most
sophisticated system available for fusing dissimilar cells. The
system is available only under license directly from Cellectis.
Please note the following important information about the
system:
1. The System must remain in the possession of the licensee.
3. The System shall only be used as described in this User
Manual.
4. Certain concepts and processes are covered by BTX patents
and patents pending.
5. BTX Cytofusion Medium C is a required component of the
Hybrimune system. Other ionic medium should not be used;
it will generate significant heating and kill cells.
6. The chamber must be cleaned after each use to prevent
ionic build up which will cause heating, kill cells, and reduce
efficiency. Please refer to chamber cleaning procedure,
page 27.
The Hybrimune™ Voltage Waveform
Generator
The waveform generator produces the voltage waveforms
programmed by the user. The desired waveform parameters and
sequences are set by the user in the user-interface application
software. When the Start button is clicked, the software is
downloaded from the computer to the microprocessor in the
waveform generator. The waveform is generated and applied to
the Electrofusion Chamber producing an electric field. Depending
on the user-input instruction, this microprocessor-controlled
waveform generator will generate:
•
Pre-Fusion Pulse AC Wave:
a low voltage,
megahertz frequency sinusoidal alternating
voltage that results in an alternating electric
field that aligns and compresses the cells in a
process called dielectrophoresis
•
Fusion Pulse:
a high voltage fusion pulse that
forms pathways in the cell membranes that
results in fusion
•
Post-Fusion Pulse AC Wave:
a second
sinusoidal alternating electric field that holds
the fused cells together while they mature
A detailed description of the waveforms is presented in page 37.
The parameter values available are listed on page 42. The User-
Interface software provides the user with the ability to fine-tune
each of the three parts of the waveform. The instructions on how
to program the waveforms may be found on page 18.
Figure 1: Hybrimune™ System
Hybrimune
™
System Components & Set-Up