selfmade vs custom antibody

CARLOS DUARTE Carlos.Duarte at CIGB.EDU.CU
Fri Dec 18 09:40:38 EST 1998


     The current trend is to prepare the antigen using
the MAP system.  I've seen variable results with this
but at least you don't have to worry about reactivity
with the carrier protein.
     If you want to use a traditional carrier such as
KLH or lysozyme then avoid carbodiimide or glutaraldehyde
as these tie up all your nice immunogenic/antigenic
lysines.  The best is some form of directional coupling
eg. with MBS and cysteine.  You can do this yourself
quite easily.


Just a comment on this subject. Even with MBS we had had some
nasty experience with conjugate peptides immunogens because in some 
cases practically all antibodies have been directed towards the MBS 
molecule itself and not against the peptide. Even when we used an imunogenic peptide 
from the V3 loop of HIV-1.  MPS  (Maleimido propionic acid 
sulccinimidyl ester) is a best option because is not as immunogenic 
as MBS. 
I agree that MAP is the best option but you must linked  your sequence 
to a known T helper cell epitope. (for example from TT) to guarantee
T helper cell involvement in the response.  
Hope it helps

Carlos A. Duarte
Centro de Ingenieria Genetica y Biotecnologia 
Division de Vacunas
Apdo 6162, Ciudad Habana CP10600
Cuba
Tel 53-7-218008
Fax 53-7-218070-336008



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