>> Have several momoclonals to a purified protein. Is there a simple means of determining if they are directed to the same or different epitopes?
>
One very simple method I have successfully used goes like this.
When a mixture of antibodies is incubated with an antigen, if the antibodies
compete against each other, the OD reading is usually the sum
of the individual ODs you obtain with identical amount of antigen and
antibody. When you do them side by side and in replicates at couple of
dilutions, you can identify if a specific pair of abys compete with a very
high degree of confidence. Although it is possible to quantitate these
values as additivity indices, I am not sure how valuable are these figures.
Please check Am J. Veterinary Research 50:1145-1149 (1989)
and J Immunol Methods 60:351-358 (1983) if you need full details
Raj Shankarappa
bsh at med.pitt.edu
