Immunoaffinity Purification and Quantification of Antibody-Toxin Conjugates

  • Edward J. Wawrzynczak
  • Alan J. Cumber
Part of the Methods in Molecular Biology book series (MIMB, volume 10)


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).


High Performance Liquid Chromatography High Affinity Binding Site High Performance Liquid Chromatography Column Elution Procedure Unconjugated Antibody 
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Copyright information

© The Humana Press, Inc., Totowa, NJ 1992

Authors and Affiliations

  • Edward J. Wawrzynczak
    • 1
  • Alan J. Cumber
    • 1
  1. 1.Drug Targeting LaboratoryInstitute of Cancer ResearchSuttonUK

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