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Updated: Jul 6, 2026
Homogeneous Time-resolved Förster Resonance Energy Transfer-based Assay for Detection of Insulin Secretion
Published on: May 10, 2018
[Establishment of the magnetic separation enzyme immunofluorescence method for detecting insulin]
Jing Xie1, Zhen-shi Liu, Shan Hu
1College of Life Science and Technology, Huazhong Science and Technology University, Wuhan 430070, China. jxmary@163.com
Aim:
To establish a convenient and sensitive magnetic separation enzyme immunofluorescence (MEIF) method for detecting human insulin.
Methods:
Two monoclonal antibodies (mAbs) were conjugated with FITC and alkaline phosphatase (AP) respectively, which were incorporated magnetic solid phase separation. Magnetic beads were coupled with sheep anti-FITC antibody as solid phase and 4-Methylumbelliferyl-phosphoric acid (4-MUP) was used as substrate to set up MElF for detecting insulin.
Results:
The sensitivity of this assay was 2.0 microIu/mL, the linear range was from 0 microIu/mL to 188.52 microIu/mL, and the intra-assay variation and inter-assay variation were 4.3%-5.2% and 2.6%-9.5%, respectively. The recovery rate of dilution was 92.6%-117% and the recovery rate of accession was 106%-121%.The result of the assay correlated well with that of magnetic enzyme chemiluminescence immunoassay system.
Conclusion:
The MEIF for detecting insulin is low at cost, sensitive, specific and stable, which can be widely used in clinical immune detection.
