Antibodies that bind M13

Lawrence P. Casson lpcasson at phoenix.Princeton.EDU
Thu May 20 04:32:26 EST 1993


I am currently attempting to make murine monoclonal antibodies against
M13.  I am doing this because I need a screening reagent that doesn't
cross react with antibodies from other sources.  For example, I have a
commercially produced sheep anti-M13 antibody (polyclonal) that cross
reacts with rabbit and goat reagents which would otherwise be useful.

I started with the quick and dirty approach which was to purify M13 by
repeated PEG precipitations and use this material to immunize mice.
The fact that other material besides M13 was present was apparent from
the physiological reaction of the mice, but fortunately, they survived.

In any case, the hybrids I have tested so far in a somewhat dirty screen
appear to react with something present in LB media in which bacteria
have been grown and removed by centrifugation.  (I coated this on
microtiter plates as a negative control to test hybrids that were
positive in a screen where bacterial supernatants containing M13 had
been used to coat plates.)  I am rescreening these on plates to which
phage-fab should be present since it is attached by a capture antibody
that binds the fab portion.

Now for the questions:

Is there a protocol for purifying M13 on a gradient as is done for phage
lambda?

Has anyone had any success in isolating monoclonal anti-M13 antibodies?

Is anyone willing to donate a hybridoma line to help me get my degree?

Thanks for any info.

Larry



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