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Custom monoclonal antibodies

Apr 1st 2015 at 12:33 AM

Hybridoma Technology
There are essentially two stages in the production
of mAbs: a) the induction of antibodyproducing
lymphoid cells in vivo and the
selection of antibody-producing hybridoma
cells in vitro; and b) the in vitro/in vivo propagation
of selected hybridoma clones. The
first stage, the formation and selection of the
hybridoma clone, involves the use of one or
more animals (except in rare cases when a
human mAb is being developed), and is carried
out in the following way:
1. The antigen is injected into mice (or
rats). The antigen is often injected in
combination with an adjuvant, to
enhance the immune response, even
though the use of adjuvant generally
leads to severe side-effects.
2. After an appropriate interval (5ñ21 days),
the immunised animals are killed and
lymphoid cells (including progenitor antibody-producing
cells) are isolated from
the spleen.
3. The lymphoid cells are fused with myeloma
cells which have been grown in vitro.
4. The two original cell types and the
newly formed hybrids are cultured in a
selective medium, such as HAT (hypoxanthine/
aminopterin/thymine) medium,
which only allows the hybridoma cells to
grow.
5. The supernatant media from the numerous
in vitro microcultures exhibiting a
recognisable growth of hybridomas are
screened for secretion of the desired antibody,
by using various immunoassay procedures.

6. The selected cells are subcultured in
vitro, using special cloning procedures to
ensure that each in vitro culture consists
of hybridomas with a single antibody
specificity only.
7. Hybridoma cells can be cryopreserved at
this stage.

 

 

 

Related Tags : monoclonal antibody production , custom monoclonal antibodies

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