Colorimetric assay of heparin in plasma based on the inhibition of oxidase-like activity of citrate-capped platinum nanoparticles

Biosens Bioelectron. 2017 Jun 15:92:442-448. doi: 10.1016/j.bios.2016.10.082. Epub 2016 Oct 30.

Abstract

We report citrate-capped platinum nanoparticles (Pt NPs) as oxidase mimetics for effectively catalyzing the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB), 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid), dopamine, and methylene blue in the presence of O2. To confirm oxidase-like activity of citrate-capped Pt NPs, their activity toward oxygen reduction reaction was studied using cyclic voltammetry and rotating ring-disk electrode method. The results obtained showed that Pt NP NPs can catalyze the oxidation of organic substrates to the colored product and the reduction of oxygen to water through a four-electron exchange process. Because the aggregation of Pt NPs can inhibit their oxidase-like activity and protamine can recognize heparin, we prepared the protamine-modified Pt NPs through direct adsorption on the surface of citrate-capped Pt NPs. The electrostatic attraction between heparin and protamine-stabilized Pt NPs induced nanoparticle aggregation, inhibiting their catalytic activity. Therefore, the lowest detectable heparin concentrations through UV-vis absorption and by the naked eye were estimated to be 0.3 and 60nM, respectively. Moreover, the proposed system enabled the determination of the therapeutic heparin concentration in a single drop of blood.

Keywords: Heparin; Oxidase mimic; Platinum nanoparticles; Protamine.

Publication types

  • Evaluation Study

MeSH terms

  • Adult
  • Anticoagulants / blood*
  • Benzidines / chemistry
  • Biomimetic Materials / chemistry
  • Biosensing Techniques / methods
  • Catalysis
  • Citric Acid / chemistry*
  • Colorimetry / methods*
  • Heparin / blood*
  • Humans
  • Limit of Detection
  • Male
  • Metal Nanoparticles / chemistry*
  • Metal Nanoparticles / ultrastructure
  • Oxidation-Reduction
  • Oxidoreductases / chemistry
  • Oxygen / chemistry
  • Platinum / chemistry*
  • Protamines / chemistry
  • Young Adult

Substances

  • Anticoagulants
  • Benzidines
  • Protamines
  • Citric Acid
  • 3,3',5,5'-tetramethylbenzidine
  • Platinum
  • Heparin
  • Oxidoreductases
  • Oxygen