title: [Establishment of the magnetic separation enzyme immunofluorescence method for detecting insulin].
abstract: 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.
authors.affiliation: College of Life Science and Technology, Huazhong Science and Technology University, Wuhan 430070, China. jxmary@163.com
authors.forename: Jing
authors.initials: J
authors.lastname: Xie
authors.forename: Zhen-shi
authors.initials: ZS
authors.lastname: Liu
authors.forename: Shan
authors.initials: S
authors.lastname: Hu
authors.forename: Ru-xiu
authors.initials: RX
authors.lastname: Cai
authors.forename: Hai
authors.initials: H
authors.lastname: Yang
authors.forename: Xiang-liang
authors.initials: XL
authors.lastname: Yang
mesh_terms: 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
chemical_list: D000911:Antibodies, Monoclonal; D007328:Insulin; D000469:Alkaline Phosphatase; D016650:Fluorescein-5-isothiocyanate