Surface functionalization of polyketal microparticles with nitrilotriacetic acid-nickel complexes for efficient protein capture and delivery

Biomaterials. 2010 Jun;31(18):4987-94. doi: 10.1016/j.biomaterials.2010.02.063. Epub 2010 Mar 25.

Abstract

Microparticle drug delivery systems have been used for over 20 years to deliver a variety of drugs and therapeutics. However, effective microencapsulation of proteins has been limited by low encapsulation efficiencies, large required amounts of protein, and risk of protein denaturation. In this work, we have adapted a widely used immobilized metal affinity protein purification strategy to non-covalently attach proteins to the surface of microparticles. Polyketal microparticles were surface modified with nitrilotriacetic acid-nickel complexes which have a high affinity for sequential histidine tags on proteins. We demonstrate that this high affinity interaction can efficiently capture proteins from dilute solutions with little risk of protein denaturation. Proteins that bound to the Ni-NTA complex retain activity and can diffuse away from the microparticles to activate cells from a distance. In addition, this surface modification can also be used for microparticle targeting by tethering cell-specific ligands to the surface of the particles, using VE-Cadherin and endothelial cells as a model. In summary, we show that immobilized metal affinity strategies have the potential to improve targeting and protein delivery via degradable polymer microparticles.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cell Line
  • Endothelial Cells / cytology
  • Endothelial Cells / metabolism
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Macrophages / metabolism
  • Mice
  • Microspheres*
  • Mitochondria / metabolism
  • Muscle Cells / metabolism
  • Nitrilotriacetic Acid / analogs & derivatives*
  • Nitrilotriacetic Acid / chemistry
  • Nitrilotriacetic Acid / metabolism
  • Organometallic Compounds / chemistry*
  • Organometallic Compounds / metabolism
  • Protein Binding
  • Surface Properties
  • Vascular Endothelial Growth Factor A / administration & dosage*

Substances

  • Organometallic Compounds
  • Vascular Endothelial Growth Factor A
  • Green Fluorescent Proteins
  • nickel nitrilotriacetic acid
  • Nitrilotriacetic Acid