I am trying to conjugate an antibody, but I want to know what the molecular structure of antibody is to see where the free thiols are and where lysine residues are for my reaction.
Also, on the same note if I want to do thiol-maleimide reaction, and I reduce the antibody to expose more thiol groups, what prevents the antibody from falling apart as the disulfide bridges are broken? I want to attach a drug to antibody without comprosing the antibody and without affecting the binding site.
There aren't supposed to be any free thiols. You can have them for 2 reasons: Your variable region has them, or some of your disulfide bonds didn't get assembled correctly. It's not usually a great idea to stick something to those, because it can be destabilizing, and the yield is never great (you can sometimes get away with it, if you must). Yes, if you reduce them on purpose, your antibody will fall apart and become inactive.
Better use the lysines or Protein A.
Arent a lot of antibody conjugated made that way though? For example, triveldy?
If i go for amine targerting, its just there are many in the variable region as well, thus interfering with its function.
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