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Hybrimune Hybridoma Production System
Publication 015-1010191 Rev 4.0 • www.btxonline.com
The Large-Volume Fusion Chamber
The Hybrimune Waveform Generator connects to a coaxial
large-volume fusion chamber. The chamber produces the various
electric fields required in the hybridoma production process.
These chambers were very carefully designed to provide optimum
alignment forces and intense electric fields necessary for efficient
electrofusion processes. When the alternating sinusoidal voltage
is applied to the inner and outer electrode rings of the chamber, a
highly non-uniform electric field is established that causes the cells
to align into ”pearl chain” formations.
Two types of chambers are provided. The first is the 2 ml chamber
(left image in Figure 2) that holds a maximum volume of 2 ml. This
chamber permits light to enter in from the bottom so that the
cell fusion process can be observed with a microscope. The 2 ml
chamber is used for process optimization. The second chamber is
the 9 ml chamber (right image in Figure 2) that holds a maximum
volume of 9 ml. This chamber is used for large batch production.
The electrical characteristics of the two chambers are equivalent.
The electrodes are coated with MEDCOAT 2000™, a chromium
coating by Electrolyzing Corporation of Ohio.
Cytofusion
®
Low Conductivity
Medium
BTX has developed a proprietary formula cell fusion medium
over several years that has under gone extensive testing. The
conductivity of this medium is low in order to minimize electrical
current flow during the application of the intense alternating
electric fields generated by the Waveform Generator. By limiting
current flow, aqueous heating is minimized which improves fusion
efficiency and viability. The medium is shown in Figure 3.
The Hybrimune Waveform Generator and the large-volume
fusion chamber were designed to work with low conductivity cell
suspensions. Do not use highly ionic buffers in the chambers. If
ionic buffers are used, two catastrophic problems will occur. First,
the cell suspension will rapidly heat causing cell death. Second,
significant convection currents will be generated in the suspension
that will inhibit cell alignment.
WARNING: ONLY CYTOFUSION® MEDIUM MAY BE USED IN
THE LARGE-VOLUME FUSION CHAMBER CHAMBERS!
Figure 2: Large-Volume Fusion Chamber
Figure 3: Cytofusion Medium C
Hybrimune
™
System Components & Set-Up