Microvolumetric determination of thrombomodulin based on competitive immunoreaction using a portable glucometer.

ABSTRACT

A new method of reducing the amount of reagent and sample for determination of thrombomodulin (TM) was developed based on competitive immunoreaction using a portable glucometer (PGM). Two types of nanocomposites, TM protein-modified magnetic nanoparticles (MNPs-TM) and TM antibody-/glucose oxidase-modified gold nanoparticles (Ab-GNPs-GOx), were prepared. Their binding product, MNPs-TM-Ab-GNPs-GOx, in the microvolumetric solution was used to catalyze the oxidation of glucose, leading to a decline of the glucose content. The TM-involved competitive immunoreaction had a negative effect on the generation of MNPs-/GNPs-based nanocomposites and inhibited the catalytic oxidation of glucose. The glucose content difference in the microvolumetric solution, which was revealed by a PGM, was in proportion to the logarithm of the TM concentration from 25 ng mL-1 to 2.5 µg mL-1. The limit of detection was 5.7 ng mL-1. Microvolumetric solution and a PGM were used in the measurement, which overcame some deficiencies of classical methods in chemo/biosensing, for example, special instrument, complicated measurement procedure, and high cost.