Pharmacia Recomb. phage Ab kit (ScFv)

Morrison Lab fergusb at microbio.lifesci.ucla.edu
Thu Nov 2 20:53:59 EST 1995



dear guys,

I just got back from a meeting in San Francisco on antibody engineering
where there was a big discussion on phage technology by most of the people
very good at it. this kit was discussed in some depth and a lot of people
said they had bad results with it - other people - a very few - said it
did actually but the ket comment was that the instructions had to be
followed exactly and to the latter i.e. no fiddling around at all. It was
also apparent that this was by no means the best library out there -
otheres are available, particularly the Nissim library from Greg winter's
lab. in cambridge, UK.
Hard to know what to see - if it's working for you then good but that is
the situation.
Needless to say, my personal opinion only

Fergus
UCLA

In article <476vhq$tri at lugb.latrobe.edu.au>, Mick Foley
<m.foley at latrobe.edu.au> wrote:

> 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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