Why polyclonal antibody from mice????

engelbert_buxbaum4 at my-deja.com engelbert_buxbaum4 at my-deja.com
Tue Nov 7 01:13:53 EST 2000


In article <3302019.973505905961.JavaMail.imail at spike.excite.com>,
  Duriya at excite.com ("Chantasingh D.") wrote:
>
> Hi All
> I need your help. I wish to know the advantage of production of
polyclonal
> antibody in mice. I used to produce polyclonal antibody from mice but
it did
> not produce ascites fluid. So I want to know can I change to produce
from
> rabbit???


Ascites is produced from cancer cells in the body cavity. Such cancer
cells can be hybridoma cells produced from B-cells (which produce the
antibody) with congenetic lymphoma cells (which donate their immortality
 to the hybridoma). Until a couple of years ago it was the preferred
method for producing monoclonal antibodies to inject the hybridomas into
the body cavity of mice and let them grow there until ascites had
accumulated in the body cavity which contained the antibody. Because
today we have incubators ('glass mouse')that can produce monoclonals
cheaply this procedure is now banned for animal protection reasons in
most countries (as you can imagine, it is rather unpleasant for the
animal to have an ascites tumor twice its normal body weight).

Injection of antigens into a mouse does not lead to ascites, but to the
production of polyclonal antibodies in the serum of the animal. Unless
you intend to use only very small amounts of antibody, or if you plan to
make monoclonals, the use of bigger animals (like rabbit, sheep or
horse, depending on your needs) is advisable, as you can draw more blood
from them. The blood is allowed to clott over night in the cold room
(i.e. do not use anticoagulants!), the clear liquid is then decanted and
saved for assays. IgG can be isolated by ammonium sulfat precipitation
and ion exchange chromatography, or by affinity chromatography.

However, before you continue working with animals, you should consult
the animal wellfare officer of your institution (this is actually a
legal requirement), and make yourself knowledgeable in the procedures
involved. A good source for reading is the 'Antibody Laboratory Manual'
by Harlow and Lane.


Sent via Deja.com http://www.deja.com/
Before you buy.






More information about the Methods mailing list