Synthesis of Maleimide-Functionalyzed HPMA-copolymers and in Vitro Characterization of the aRAGE- And Human Immunoglobulin (huIgG)-polymer Conjugates

Macromol Biosci. 2013 Feb;13(2):203-14. doi: 10.1002/mabi.201200344. Epub 2012 Dec 13.

Abstract

Herein the synthesis of antibody-polymer conjugates, with a quite narrow dispersity based on the polymer HPMA, are reported. These conjugates are synthesized by coupling antibodies to maleimide-functionalized poly(N-(2-hydroxypropyl)-methacrylamide) (poly-HPMA) copolymers derived through reversible addition-fragmentation chain transfer (RAFT) polymerization of pentafluorophenyl methacrylate via the intermediate step of an activated ester polymer. We develop a protocol that allows the attachment of two different model antibodies, monoclonal anti-RAGE (receptor for advanced glycation end-products) antibody, and polyclonal human immunoglobulin (huIgG). Modification of the antibody and conjugation is monitored by SDS-PAGE electrophoresis. Preserved affinity is demonstrated by Western Blott and cell-uptake analysis, for example, to cells of the immune system.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / chemistry*
  • Binding Sites
  • Cell Line
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Immunoglobulins / chemistry*
  • Immunoglobulins / metabolism
  • Maleimides / chemistry*
  • Polymethacrylic Acids / chemical synthesis
  • Polymethacrylic Acids / chemistry*
  • Receptor for Advanced Glycation End Products
  • Receptors, Immunologic / immunology*
  • Receptors, Immunologic / metabolism

Substances

  • Antibodies, Monoclonal
  • Immunoglobulins
  • Maleimides
  • Polymethacrylic Acids
  • Receptor for Advanced Glycation End Products
  • Receptors, Immunologic
  • maleimide
  • Duxon