Homogeneous bispecifics by disulfide bridging

Bioconjug Chem. 2014 Aug 20;25(8):1395-401. doi: 10.1021/bc5002467. Epub 2014 Aug 1.


We report on a chemical platform to generate site-specific, homogeneous, antibody-antibody conjugates by targeting and bridging disulfide bonds. A bispecific antibody construct was produced in good yield through simple reduction and bridging of antibody fragment disulfide bonds, using a readily synthesized bis-dibromomaleimide cross-linker. Binding activity of antibodies was maintained, and in vitro binding of target antigens was observed. This technology is demonstrated through linking scFv and Fab antibody fragments, showing its potential for the construction of a diverse range of bispecifics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibody Specificity*
  • Disulfides / chemistry*
  • Protein Multimerization
  • Protein Structure, Quaternary
  • Single-Chain Antibodies / chemistry*
  • Single-Chain Antibodies / immunology*
  • Substrate Specificity


  • Disulfides
  • Single-Chain Antibodies