Blood-brain barrier molecular trojan horse enables imaging of brain uptake of radioiodinated recombinant protein in the rhesus monkey

Bioconjug Chem. 2013 Oct 16;24(10):1741-9. doi: 10.1021/bc400319d. Epub 2013 Oct 3.


Recombinant proteins are large molecule drugs that do not cross the blood-brain barrier (BBB). However, BBB-penetration of protein therapeutics is enabled by re-engineering the recombinant protein as IgG fusion proteins. The IgG domain is a monoclonal antibody (mAb) against an endogenous BBB receptor-mediated transport system, such as the human insulin receptor (HIR), and acts as a molecular Trojan horse to ferry the fused protein across the BBB. In the present study, a recombinant lysosomal enzyme, iduronate 2-sulfatase (IDS), is fused to the HIRMAb, and BBB penetration of the IDS alone vs the HIRMAb-IDS fusion protein is compared in the Rhesus monkey. Recombinant IDS and the HIRMAb-IDS fusion protein were radiolabeled with indirect iodination with the [(125)I]-Bolton-Hunter reagent and with direct iodination with Iodogen/[(125)I]-idodine. IDS and the HIRMAb-IDS fusion protein have comparable plasma pharmacokinetics and uptake by peripheral organs. IDS does not cross the BBB. The HIRMAb-IDS fusion protein crosses the BBB and the brain uptake is 1% of injected dose/brain. Brain imaging shows HIRMAb-IDS penetration to all parts of brain, and immunoprecipitation of brain radioactivity shows intact fusion protein in brain. The use of BBB molecular Trojan horses enables brain imaging of recombinant proteins that are re-engineered for BBB transport.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / analysis
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / pharmacokinetics*
  • Blood-Brain Barrier / diagnostic imaging
  • Blood-Brain Barrier / metabolism*
  • Brain / diagnostic imaging
  • Brain / metabolism
  • Horses
  • Humans
  • Iduronate Sulfatase / analysis
  • Iduronate Sulfatase / pharmacokinetics*
  • Iodine Radioisotopes / analysis
  • Macaca mulatta
  • Radiography
  • Receptor, Insulin / immunology
  • Recombinant Fusion Proteins / analysis
  • Recombinant Fusion Proteins / immunology
  • Recombinant Fusion Proteins / pharmacokinetics*
  • Succinimides / analysis


  • Antibodies, Monoclonal
  • Iodine Radioisotopes
  • Recombinant Fusion Proteins
  • Succinimides
  • Bolton-Hunter reagent
  • Receptor, Insulin
  • Iduronate Sulfatase