Pharmacia Recomb. phage Ab kit (ScFv)

Mick Foley m.foley at latrobe.edu.au
Wed Nov 1 00:11:22 EST 1995


Deborah and Rajendra,

I was interested in your comments on the recombinant phage antibody 
diplay system from Pharmacis. We have been using this system to clone 
antibody genes from immunised mice. After each step of panning we do PCRs 
from the pools of phage and from individual colonies, using primers that 
span the MCS. We find that we get three bands! The correct size (900bp) a 
450bp - presumably some deletion product and possibly the same as you 
guys see and also a 1.5 Kb band which we have not got a clue as to what 
it is. We are at the moment sequencing this band. Individual clones show 
one or other of these bands but each clone is only one band. I agree that 
you have to fiddle around with this system, but for immune spleens it 
does seem to work. We have isolated scFv clones which bind to the antigen 
by Elisa and dot blots from a library of just over 20,000 clones 
(although this is probably dependat on the immunization procol)
It's good to see the comments of others using it.
Cheers, 
        Mick




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