Carboxylated magnetic microbead-assisted fluoroimmunoassay for early biomarkers of acute myocardial infarction.
A carboxylated superparamagnetic microbead-assisted sandwich fluoroimmunoassay was successfully demonstrated for the analysis of the early protein markers, myoglobin and human heart-type fatty acid-binding protein (H-FABP), associated with acute myocardial infarction. This assay approach consisted of the preparation of superparamagnetic polymer microbeads using a dispersion polymerization, followed by grafting of capture antibodies (monoclonal anti-H-FABP 10E1 and anti-myoglobin 7C3) onto the polymer microbeads using EDC-NHS protocol, and then a sequential sandwich fluoroimmunoassay using detection antibodies (FITC-labeled anti-H-FABP 9F3 and FITC-labeled anti-myoglobin 4E2). The Fe(3)O(4) nanoparticles and carboxylated Fe(3)O(4)-polymer microbeads were characterized by scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectrophotometry, vibrating sample magnetometry, and X-ray diffraction. The fluoroimmunoassay images were recorded using a confocal laser-scanning microscope, and the average fluorescence intensity of the microbeads was found to correspond to the concentration of each cardiac marker, in agreement with the results obtained by a spectrofluorophotometer. The carboxylated magnetic microbead-assisted protocol could be utilized to semi-quantitatively detect both myoglobin and H-FABP.
Wang
Jinyi
J
College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, PR China. jywang@nwsuaf.edu.cn
Wang
Qiang
Q
Ren
Li
L
Wang
Xueqin
X
Wan
Zongfang
Z
Liu
Wenming
W
Li
Li
L
Zhao
Huiying
H
Li
Manlin
M
Tong
Dewen
D
Xu
Juan
J
D000179:Acrylates / Q000737:chemistry; D000208:Acute Disease; D000906:Antibodies; D015415:Biomarkers / Q000032:analysis; D002264:Carboxylic Acids / Q000378:metabolism*; D005290:Ferric Compounds / Q000378:metabolism; D005453:Fluorescence; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D008280:Magnetics; D008863:Microspheres; D009203:Myocardial Infarction / Q000175:diagnosis*; D053758:Nanoparticles / Q000648:ultrastructure; D011137:Polystyrenes / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared; D013997:Time Factors; D014961:X-Ray Diffraction
D000179:Acrylates; D000906:Antibodies; D015415:Biomarkers; D002264:Carboxylic Acids; D005290:Ferric Compounds; D011137:Polystyrenes; C000499:ferric oxide; C036658:acrylic acid