[Establishment of the magnetic separation enzyme immunofluorescence method for detecting insulin].
OBJECTIVE
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.


CONCLUSIONS
The MEIF for detecting insulin is low at cost, sensitive, specific and stable, which can be widely used in clinical immune detection.
Xie
Jing
J
College of Life Science and Technology, Huazhong Science and Technology University, Wuhan 430070, China. jxmary@163.com
Liu
Zhen-shi
ZS
Hu
Shan
S
Cai
Ru-xiu
RX
Yang
Hai
H
Yang
Xiang-liang
XL
D000469:Alkaline Phosphatase / Q000737:chemistry*; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry*; D005455:Fluorescent Antibody Technique / Q000379:methods*; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D018189:Immunomagnetic Separation / Q000379:methods*; D007328:Insulin / Q000032:analysis*; D015203:Reproducibility of Results
D000911:Antibodies, Monoclonal; D007328:Insulin; D000469:Alkaline Phosphatase; D016650:Fluorescein-5-isothiocyanate