> From: jcc at alumnae.caltech.edu (jc catalano)
>> Subject: how to get monoclonal antibodies directed against mouse proteins
>> Date: 11 Jan 1996 08:18:50 GMT
>> Hello-
>> This may sound somewhat silly, but suppose that I want to study a
> protein
> from a mouse and would like monoclonal antibodies, is there a
> convenient
> way to do this?
> Being a non-immunologist, I understand that the usual way to make
> mAB's is
> to inject mice with protein X, then take spleen cells that have
> antibodies
> directed against X, fuse with myeloma cells, and pick out the
> monoclonals
> via an ELISA.
>> So do you use a rat or another animal to produce anti-mouse
> monoclonals?
>> Any help or references appreciated. Thanks in advance.
>> j.c.
The strategy we now adopt for almost all of our poly- and monoclonal
antibody production is to first immunize sheep. They have the advantages
of being long lived, large (lots of serum) and cheep to keep (outside
munching grass). We screen the polyclonal antibodies we get from the
sheep, and if the polyclonal antibody is good enough for the proposed use
(immunoassay, immunoaffinity, blotting etc.) we bank a few hundred mL,
use it, and move on to the next problem (oh, and put the sheep on a
pension - just in case).
If however we do actually need a monoclonal for whatever reason (we need
HUGE amounts or there are too many x-reactivities in the polyclonal which
can not be easily stripped-out of the serum - we then produce a
ovine/murine hetero-hybridoma from peripheral lymphocytes harvested from
the sheep (and the sheep still gets to go on a pension).
In our laboratory this twin-track approach works well. The ovine
monoclonals have very high affinities, and in clinical immunoassay
applications no HAMA effects.
Hope this helps, it is one of the ways you can overcome your problem.
Pete
