Superparamagnetic microsphere-assisted fluoroimmunoassay for rapid assessment of acute myocardial infarction

作者：Wang Jinyi^*; Ren Li; Wang Xueqin; Wang Qiang; Wan Zongfang; Li Li; Liu Wenming; Wang Xuming; Li Manlin; Tong Dewen; Liu Ajing; Shang Bingbing

来源：Biosensors and Bioelectronics, 2009, 24(10): 3097-3102.

DOI：10.1016/j.bios.2009.03.033

• Superparamagnetic poly(styrene-divinylbenzene-acrylamide) microsphere
• Myoglobin
• Human heart-type fatty acid binding protein
• Fluoroimmunoassay

Rapid assessment of acute myocardial infarction (AMI) was successfully demonstrated using an improved superparamagnetic polymer microsphere-assisted sandwich fluoroimmunoassay to detect two early cardiac markers-myoglobin and human heart-type fatty acid binding protein (H-FABP). This assay used a preparation of superparamagnetic poly(styrene-divinylbenzene-acrylamide) microspheres, glutaraldehyde-coupled capture antibodies (monoclonal anti-myoglobin 7C3 and anti-H-FABP 10E1) grafted onto the polymer microspheres, and a sequential sandwich fluoroimmunoassay using detection antibodies (FITC-labeled anti-myoglobin 4F2 and FITC-labeled anti-H-FABP 9F3). Characterization of the polymer microspheres by TEM, SEM and Fourier transform infrared spectroscopy (FT-IR) showed that the microspheres were uniformly round with an average diameter of 4.12 mu m, and had a Fe(3)O(4)-polymer core-shell structure (shell thickness was about 84 nm) with 0.22 mmol/g amino groups on their surfaces. The magnetic behavior of the Fe(3)O(4)-polymer microspheres was superparamagnetic (M(s) = 13 emu/g, H(c) = 13.1 Oe). Fluorescence images of the post-immunoassay microspheres recorded using a confocal laser-scanning microscope showed that the average fluorescence intensity was correlated with the concentration of cardiac markers, in agreement with the results obtained by an F-4500 FLspectrophotometer: this indicated that the fluoroimmunoassay could be used to semi-quantitatively detect both myoglobin and H-FABP. The detection limit was 25 ng/mL for rnyoglobin and 1 ng/mL for H-FABP.

• 出版日期2009-6-15
• 单位西北农林科技大学