Technology

Cyto Pulse antibody discovery technology enables efficient creation of a large variety of monoclonal antibodies.  Monoclonal antibodies are increasingly being approved for clinical applications and the need for monoclonal antibodies for clinical diagnostics is strong.  Phases in the discovery and creation of monoclonal antibodies are: target selection, vaccination of animals (usually mice or transgenic mice), immortalization of B cells or cloning of antibody genes and manufacturing of the antibody.  After target selection, some paths to antibody product are:

Cyto Pulse technology begins at vaccination and continues through production.  For the first step, vaccination of animals, DNA vaccination is recommended (OncoVet™).  Advantages of DNA vaccination over protein based vaccination for activated B cell production include a vaccine with a defined specificity, expression of protein in its native conformation, induction of high avidity antibodies, and the minimization of labor, bypassing the need to produce protein in vitro.  In some cases, the protein is very difficult to produce in their native configurations.

After DNA vaccination, B-cells may be immortalized to express their antibodies. Cyto Pulse’s electrofusion technology (Hybrimune™) was designed for this task. Patented electrofusion chamber design allows large volume electrofusion for hybridoma production. Specialized waveforms permit efficient and gentle electrofusion.

Alternatively, immunoglobulin gene can be cloned and expressed in cultured cells such as CHO. Cyto Pulse's large volume transfection technology (CytoLVT-S™) is designed to transiently transfect cells in sufficient quantities for seeding bioreactors. Antibodies expressed from the transfected cells are harvested directly from the bioreactor.

Systems Used in Antibody Discovery

OncoVet™
OncoVet™ delivers DNA vaccines to laboratory animals by electroporation. OncoVet™ has several unique features. OncoVet™ delivers vaccines into the skin, rather than the muscle. Skin delivery is convenient and more immunologically active than muscle. OncoVet™ also utilizes Cyto Pulse's patented PulseAgile® technology for pulse delivery. Digitally controlled PulseAgile® protocols electroporate delivery sites at a rapid rate (10 pulses in under 0.3 seconds) and produce robust B and T cell responses.

Aldevron has licensed Cyto Pulse OncoVet™ technology and is available to conduct evaluation studies and projects.

 

Hybrimune™
The electrofusion system, Hybrimune™, also has some unique features.  The most visible is that the large processing capacity.  Up to 9 ml of cell mixture can be processed at each electrofusion.  This compares to the few hundred microliters that could be processed prior to the introduction of Hybrimune™.  High processing volume can be attributed to patented electroporation chamber configuration combined with proprietary electrofusion medium.  In addition, Hybrimune™ delivers a cell alignment waveform in a patented sequence that enhances hybridoma yield.  Compared to standard PEG fusion, yield improvements of 10 fold or more are observed.

CytoLVT-S™
CytoLVT-S™ system is a large volume transfection system that can process 2 to 20 mL.

CytoLVT-P™
CytoLVT-P™ system is a large volume transfection system that can process 50 to 1000 mL with cell density of cells at cells/mL which would seed a 100 liter bioreactor at 1 million cells/mL.

Cyto Pulse References:

Medarex
Morphotek

Development Partners
Hybrimune™	Medarex, Milpitas, CA
Cyto-LVT-S™	NovImmune, Geneva, CH
Onco Vet™	Amgen, CA Aldevron, Fargo, ND Statens Serum Institut, Copenhagen, Denmark

 

Numerous commercial hybridoma production licenses for Monoclonal Antibody discovery are in place worldwide, many users have multiple licenses:

• Medarex, Princeton, NJ, USA
• Novo Nordisk, Denmark
• Genmab, Netherlands
• Morphotek, Exton, PA, USA
• NovImmune, Geneva, Switzerland
• Schering-Plough Biopharma (formerly DNAX Research, Inc.), USA
• Genentech, Inc., San Francisco, CA, USA
• Mapp Biopharmaceutical, Inc., San Diego, CA, USA
• OncoMed Pharmaceuticals, Mountain View, CA, USA
• Amgen, Inc., USA
• Takeda Pharmaceutical, Osaka, Japan
• ZymoGenetics, Seattle, WA, USA
• Regeneron Pharmaceuticals, Inc., Tarrytown, NY, USA
• Genomics Institute of Novartis Research Foundation (GNF), San Diego, CA, USA
• Galaxy Biotech, Cupertino, CA, USA
• Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA
• Daiichi Sankyo Co., Ltd., Tokyo, Japan
• Organon Research, Schering-Plough, Cambridge, MA, USA
• Spaltudaq Corporation, Seattle, WA, USA
• Rockland Immunochemicals, Inc., Boyertown, PA, USA
• Roche Diagnostics, Penzberg, Germany
• Merck, West Point, PA, USA
• Acceleron Pharma, Inc., Cambridge, MA, USA
• GlaxoSmithKline, UK
• Others in negotiation

 

This system is available under direct license agreement with Cyto Pulse.
Please Contact Cyto Pulse for Additional Information

Part No. 016-100543