Tuning the Hydrolytic Stability of Next Generation Maleimide Cross-Linkers Enables Access to Albumin-Antibody Fragment Conjugates and tri-scFvs

Bioconjug Chem. 2018 Feb 21;29(2):486-492. doi: 10.1021/acs.bioconjchem.7b00795. Epub 2018 Jan 31.

Abstract

We describe investigations to expand the scope of next generation maleimide cross-linkers for the construction of homogeneous protein-protein conjugates. Diiodomaleimides are shown to offer the ideal properties of rapid bioconjugation with reduced hydrolysis, allowing the cross-linking of even sterically hindered systems. The optimized linkers are exploited to link human serum albumin to antibody fragments (Fab or scFv) as a prospective half-life extension platform, with retention of antigen binding and robust serum stability. Finally, a triprotein conjugate is formed, by linking scFv antibody fragments targeting carcinoembryonic antigen. This tri-scFv is shown to infer a combination of greater antigen avidity and increased in vivo half-life, representing a promising platform for antibody therapeutic development.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cross-Linking Reagents / chemistry*
  • Humans
  • Hydrolysis
  • Immunoconjugates / chemistry*
  • Immunoglobulin Fab Fragments / chemistry
  • Maleimides / chemistry*
  • Models, Molecular
  • Serum Albumin, Human / chemistry*
  • Single-Chain Antibodies / chemistry*

Substances

  • Cross-Linking Reagents
  • Immunoconjugates
  • Immunoglobulin Fab Fragments
  • Maleimides
  • Single-Chain Antibodies
  • Serum Albumin, Human