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Hybrimune Hybridoma Production System
Publication 015-1010191 Rev 4.0 • www.btxonline.com
Available Myelomas or Heteromyelomas
Note: This list is not exhaustive. It was made from literature
searches and a search of the ATCC and Coriell Institute’s web sites.
Carefully research intellectual property issues relating to these cells
before using them since some of these cell lines may be patented.
It does not indicate which cell lines are better suited as fusion
partners for the species indicated. It is meant to provide a starting
point for finding a good fusion partner.
Human
• U-226AR
• GM1500GTGAL2 (Coriell Institute, GMO4672)
• HuNS1 (ATCC CRL8644)
• SHM-D33 (ATCC CRL-1668)
• KR-12 (ATCC CRL8658): Derived from GM1500
Mouse
IgG secreting mouse myeloma cells:
• MOPC 21
• P3X63AG8 (ATCC #TIB9)
Non-antibody mouse secreting cells (some produce light chains):
• SP2/0-Ag14 (ATCC #CRL 1581)
• NS-1 (ATCC #TIB18)
• P3.X63AG8.653 (ATCC #CRL 1580)
• F0 (ATCC CRL1646)
• S194/5.XXO.BU.1 (ATCC #CRL 1580, TIB20)
• FOX-NY (ATCC #CRL 1732).
Rat
• 210. RCY3.Agl.2.3
Rabbit
• 240E1-1 (Spieker-Polet 1995)
B Cells
Preparation of B cells is more complicated. The general steps are in
vivo or in vitro immunization, collection and purification of B cells,
and optional activation of B cells. There are many ways to do this
series of procedures and all of them will not be covered.
Immunization
One immunization procedure follows. Immunize mice every 3
to 4 weeks. The first immunization can use complete Freund’s
adjuvant. Subsequent immunizations should employ other
adjuvants such as incomplete Freund’s adjuvant. Other more
modern adjuvants can be used.
Bleed the mice periodically to determine if they are producing
polyclonal antibody to the antigen of interest. Three days before
harvesting spleen cells, a final boost of antigen without adjuvant
can be given intravenously to activate B cells and localize B cells to
the spleen.
An alternate immunization procedure is to use DNA vaccines
delivered using intradermal inoculation of plasmid plus
electroporation. Mice are immunized every 3 to 4 weeks. The
immunization can be either a series of DNA vaccines or a DNA
prime with protein boost. A final boost is given 3 to 7 days prior
to harvest. For details, consult BTX regarding the AgilePulse™ DNA
vaccine delivery system.
Screening
Test Bleeds
• Identify mice with ear tags or ear clips.
• Apply grease to tail (an area of 3/4 inch, from base
toward tip).
• Use extremely sharp blades to make slice (EM blades).
Be careful not to cut through the tail but make cut large
enough to draw about 2.5 heparinized capillary tubes.
• Place blood into mouse clot tubes.
• Apply pressure to tail wound to stop the bleeding.
• Spin down the blood at max speed for 5 minutes in the
microfuge.
• Draw off and store sera at -20
o
C until you can test on
a blot or ELISA at 1:100 or at 1:50.
Evaluating sera:
Use an ELISA or Western Blot to evaluate
antibody production in mice.
Producing Fusion Products Using Hybrimune
™