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).
KeywordsHigh Performance Liquid Chromatography Filtration Glycine Electrophoresis H2SO4
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