Conjugation of anticancer drugs through endogenous monoclonal antibody cysteine residues

Methods Enzymol. 2012;502:123-38. doi: 10.1016/B978-0-12-416039-2.00006-9.

Abstract

Many methods have been described for the conjugation of drugs to monoclonal antibodies. The presence of a discrete number of readily reducible disulfides in the common IgG subtypes presents a convenient opportunity for conjugation to cysteine residues with thiol-reactive drug-linkers. Such conjugates can be prepared by a straightforward two-step reaction scheme involving the reduction of the antibody disulfides to the desired number of average thiols per antibody, followed by addition of the drug-linker, ideally with a maleimido functionality for rapid, selective reaction. In a discovery setting, this basic method can be scaled down to produce microgram quantities of conjugate for early screening, and in a manufacturing setting can be scaled up to produce grams or kilograms of conjugate for clinical trials and commercialization. The resulting conjugates are readily characterized using common HPLC methods.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / chemistry*
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / metabolism
  • Antibodies, Monoclonal / pharmacology
  • Antigens / immunology
  • Antigens / metabolism
  • Antineoplastic Agents / chemistry*
  • Antineoplastic Agents / immunology
  • Antineoplastic Agents / metabolism
  • Antineoplastic Agents / pharmacology
  • Cell Line, Tumor
  • Chromatography, High Pressure Liquid
  • Cross-Linking Reagents / chemistry
  • Cysteine / chemistry
  • Cysteine / metabolism*
  • Disulfides / chemistry
  • Disulfides / metabolism
  • Drug Carriers / chemistry
  • Drug Carriers / metabolism*
  • Drug Carriers / pharmacology
  • Ethylmaleimide / chemistry
  • Humans
  • Hybridomas / immunology
  • Hybridomas / metabolism
  • Immobilized Proteins / chemistry*
  • Immobilized Proteins / metabolism
  • Immunoconjugates / chemistry
  • Immunoconjugates / metabolism*
  • Immunoconjugates / pharmacology
  • Mice
  • Molecular Structure
  • Neoplasms / drug therapy*
  • Neoplasms / immunology
  • Neoplasms / metabolism
  • Neoplasms / pathology
  • Oxidation-Reduction
  • Sulfhydryl Reagents / chemistry

Substances

  • Antibodies, Monoclonal
  • Antigens
  • Antineoplastic Agents
  • Cross-Linking Reagents
  • Disulfides
  • Drug Carriers
  • Immobilized Proteins
  • Immunoconjugates
  • Sulfhydryl Reagents
  • Cysteine
  • Ethylmaleimide