Skip to Content
Merck
  • Establishment of Magnetic Microparticles-Assisted Time-Resolved Fluoroimmunoassay for Determinating Biomarker Models in Human Serum.

Establishment of Magnetic Microparticles-Assisted Time-Resolved Fluoroimmunoassay for Determinating Biomarker Models in Human Serum.

PloS one (2015-06-24)
Zhi-Qi Ren, Tian-Cai Liu, Si-Hui Zhuang, Guan-Feng Lin, Jing-Yuan Hou, Ying-Song Wu
ABSTRACT

In order to early screen and detect suspected biomarkers from pathogens and the human body itself, tracers or reaction strategies that can act as signal enhancers have been proposed forth at purpose. In this paper, we discussed the applicability of magnetic microparticles-assisted time-resolved fluoroimmunoassay (MMPs-TRFIA) for sensitive determination of potential analytes. Hepatitis B e antigen, antibody to hepatitis B surface antigen and free triiodothyronine were used as biomarker models to explore the reliability of the method. By coupling with bioprobes, MMPs were used as immunoassay carriers to capture target molecules. Under optimal condition, assay performance, including accuracy, precision and specificity, was outstanding and demonstrated satisfactory. To further evaluate the performance of the MMPs-TRFIA in patients, a total of 728 serum samples from hospital were analyzed for three biomarkers in parallel with the proposed method and chemiluminescence immunoassay kit commercially available. Fairly good agreements are obtained between the two methods via data analysis. Not only that but the reliability of MMPs-TRFIA has also been illustrated by three different reaction models. It is confirmed that the novel method modified with MMPs has been established and showed great potential applications in both biological detection and clinical diagnosis, including big molecule protein and low molecular weight haptens.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
MES solution, BioUltra, for molecular biology, 0.5 M in H2O
Sigma-Aldrich
N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride, BioXtra
Sigma-Aldrich
N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride, crystalline
Sigma-Aldrich
MES, low moisture content, ≥99% (titration)
Sigma-Aldrich
Monoclonal Anti-HBV antigen HBsAg antibody produced in mouse, clone HB5, purified immunoglobulin, buffered aqueous solution
Sigma-Aldrich
1,2-Dichloroethane, LR, ≥99%
Sigma-Aldrich
1,2-Dichloroethane, AR, ≥99.5%
Sigma-Aldrich
N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride, ≥99.0% (AT)
Sigma-Aldrich
N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride, commercial grade, powder
Sigma-Aldrich
1,2-Dichloroethane, anhydrous, 99.8%
Sigma-Aldrich
N-Hydroxysuccinimide, 98%