Antibody raised against an Antibody

Bernard P. Murray, PhD bpmurray*STUFFER* at
Mon Feb 15 20:29:12 EST 1999

In article <utNx2.4480$_A1.25710 at>, "8675309"
<ke8675309 at> wrote:

> I am a newcomer to the newsgroup and field of molbio so i beg forgiveness
> for my lack of vocabulary. I am wondering if and antibody(i will call it
> A1)could be raised against an antibody that has already been raised against
> the tip of a conjugative pilus (of say E.coli K12), with the resultant A1
> still retaining bioactivty such that it will associate with the same
> membrane protien that the pilus tip normally does to form a conjugative
> junction with a receptive bacteria or yeast for that matter. If this is
> possible, can a marker be associated with A1 without disturbing said
> bioactivity. Can A1 be made to irreversibly bind to said membrane protiens.
> Thankyou for your time.
> Kent Lavallie
> Undergrad UofO

In principle you can make an anti-idiotype antibody that
has a binding surface that behaves like the pilus tip.
This method has been demontrated in other circumstances
(eg. anti-idiotype antibodies raised against original
antibodies that bind to neurotransmitters can bind to
the receptor of the neurotransmitter).
     The main problem is that this normally requires a
(good) monoclonal antibody as the antigen and an
efficient screen (if I recall correctly good anti-idiotype
antibodies are quite rare).

Tagging the resultant antibody should not be a major
problem.  Conjugation of primary antibodies is fairly
routine.  You may even be able to target the region
that is labelled, eg. take protein-A labelled with
eg. peroxidase and bind this to the antibody (should
be Fc region selective) and then attach it permanently
with a cross-linking agent.

Similarly, if your antibody is good and binds well
to the antigen you can immunoprecipitate and again
cross-link and a proportion of your membrane protein
should have the antibody permanently bound.

Bottom line:  Theoretically possible, Practically laborious

     Good luck,
Bernard P. Murray, PhD
Dept. Cell. Mol. Pharmacol., UCSF, San Francisco, USA

More information about the Methods mailing list