Conjugation of von Willebrand factor-binding domain of platelet glycoprotein Ibα to size-controlled albumin microspheres

Shinji Takeoka, Yuji Teramura, Haruki Ohkawa, Yasuo Ikeda, Eishun Tsuchida

Research output: Contribution to journalArticlepeer-review

28 Citations (Scopus)

Abstract

Albumin microspheres (AMS), of which the average diameter is 240 ± 10 nm, were prepared by pH control and heat treatment. Cytochrome c and rGPIbα; a water-soluble fragment of the α chain of a recombinant platelet glycoprotein (GP)Ib containing a von Willebrand factor (vWf)-binding site were selected as a receptor protein. Cytochrome c was used as a probe protein for monitoring. Onto the surface of the AMS and those proteins, N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP) was reacted through the amide linkage to obtain PD-AMS, PD-cytochrome c, and PD-rGPIbα, respectively. The latter two were further reduced to SH-cytochrome c and SH-rGPIbα by dithiothreitol and conjugated with PD-AMS by a thiol - disulfide exchange reaction. The resulting AMS contain cytochrome c or rGPIbα of about 25000 or 2500 molecules, respectively. The addition of ristocetin to the rGPIbα-AMS in the presence of vWf caused specific aggregation. Furthermore, the rGPIbα-AMS enhanced the ristocetin induced platelet aggregation in a low platelet concentration (4.0 × 107/mL).

Original languageEnglish
Pages (from-to)290-295
Number of pages6
JournalBiomacromolecules
Volume1
Issue number2
DOIs
Publication statusPublished - 2000 Jan 1

ASJC Scopus subject areas

  • Bioengineering
  • Biomaterials
  • Polymers and Plastics
  • Materials Chemistry

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