polyclonal Ab against the variable region - (Jul/26/2006 )

Hi all.
I need help please.
I want to produce polyclonal Ab against the part of the variable region (CDR1 mostly) of human monoclonal Ab (IgG).
In my previous trials I've injected mAb into the rabbit and cleaned the pAb on G-sepharose column. Eventually I’ve got the non specific pAb population i.e. it recognizes many common IgG determinants.
I have very little quantity of the mAb so I cannot slice it in order to rid of the common parts (like Fc for example).
Please advice if there are ways to produce determinant specific anti-Ab antibodies (poly or mono).
Thanks in advance.
Pery

hi,
use CDR1 peptide for immunizations to raise polyclonal antibodies, then u can purify ur pAb using peptide affinity column.

hey, u cleaned ur pAb with G-sepharose, that means it can bind to Fc fragment of any kind of antibody hence u possibly could have purified all other types of antibodies from rabbit serum.
i hope u understood the point y u got many common antibodies which binds to other igG determinants.

in my opinion, G-sepaharose works better for purification of monoclonal antibodies from culuture supernatants since there u have only MAbs.

gud luk
payeli.

Dear Payeli,
Thanks for the fast reply.
As I wrote, my problem is that I have very small quantity of this mAb.
Therefore I cannot have it's CDR1 (not enough mAb for slicing and cleaning process).
I guess it no point of using CDR1 from other Abs either, isn't it?
Anyway, may be you know some methods that allows CDR production using small quantity of Ab?
Thanks again
Pery



[quote name='payeli' date='Jul 26 2006, 08:11 PM' post='61341']
hi,
use CDR1 peptide for immunizations to raise polyclonal antibodies, then u can purify ur pAb using peptide affinity column.

hey, u cleaned ur pAb with G-sepharose, that means it can bind to Fc fragment of any kind of antibody hence u possibly could have purified all other types of antibodies from rabbit serum.
i hope u understood the point y u got many common antibodies which binds to other igG determinants.

in my opinion, G-sepaharose works better for purification of monoclonal antibodies from culuture supernatants since there u have only MAbs.

gud luk
payeli.

hi,
i donot mean peptide from ur antibody.
i mean synthetic peptide, ofcourse it is little bit expensive but this one possibility.

regards
payeli.

Thanks