Immunoaffinity purification and quantification of antibody-toxin conjugates

Cytotoxic antibody-toxin conjugates made using antibodies and ribs some-inactivating proteins (RIPs) are prepared using chemical crosslinking methods (1,2 and this vol., Chapter 31 ). Gel permeation chromatography is used as a first step to purify conjugate molecules from the reaction mixture. This procedure removes protein aggregates, the excess of RIP employed in the conjugation reaction, and low molecular weight byproducts. However, a significant fraction of the resulting conjugate preparation consists of unconjugated antibody that cannot be completely separated from the conjugate on the basis of size discrimination alone (see chapter 31 ).