Chemiluminescence Immunoassays for Simultaneous Detection of Three Heart Disease Biomarkers Using Magnetic Carbon Composites and Three-Dimensional Microfluidic Paper-Based Device

Abstract

Chemiluminescence (CL) immunoassays for simultaneous detection of early acute myocardial infarction (AMI) biomarkers, including copeptin, heart-type fatty acid binding protein (h-FABP), and cardiac troponin I (cTnI), were developed by using Co2+/N-(aminobutyl)-N-(ethylisoluminol) (ABEI) functionalized magnetic carbon composite (Co2+-ABEI-Fe3O4@void@C) as an interface and a three-dimensional microfluidic paper-based device (3D μPAD) as a detection system. For CL immunoassays, Co2+-ABEI-Fe3O4@void@C was assembled with chitosan (CS) and gold nanoparticle-conjugated antibody (Au-Ab) sequentially to form the sensing platform (Co2+-ABEI-Fe3O4@void@C-CS/Au-Ab). In the presence of antigen (Ag), Ag was captured by the sensing interface to form an immunocomplex, leading to an increase in CL intensity due to the catalysis of −COO– existing in Ag. A 3D μPAD with three detection zones for simultaneous detection of copeptin, h-FABP, and cTnI was designed and fabricated to obtain time-resolved CL signals. Three kinds of immunocomplexes formed with copeptin, h-FABP, and cTnI were added to three detection zones of 3D μPAD, respectively. After injecting H2O2, three time-resolved CL signals were generated in one CL detection run by virtue of time-delayed transport of H2O2 to different detection zones. The three time-resolved CL signals were used for the simultaneous determination of copeptin, h-FABP, and cTnI. The detection limit of copeptin, h-FABP, and cTnI was 0.40 pg/mL, 0.32 pg/mL, and 0.50 pg/mL, respectively, which is at least 1 order of magnitude lower than most of the reported immunoassays. The immunoassays could be directly used for the detection of copeptin, h-FABP, and cTnI in human serum samples. The proposed immunoassays are simple, fast, sensitive, and selective, and are of great application potential in early diagnosis and treatment of AMI.

Application of Microfluidics in Immunoassay: Recent Advancements

“…In 2020, as shown in Figure 12 composite (Co2+-ABEI-Fe 3 O 4 @ void@C) was used as an interface. e LOD of the experiment was 0.40 pg/mL, 0.32 pg/mL, and 0.50 pg/mL, respectively [70]. In 2020, as shown in Figure 12(b), a novel magnetic microfluidic sensor with nanostar antennas was developed for the multiplexing detection of cardiac troponin I (cTnI) and neuropeptide Y (NPY) in which the LOD of 0.0055 ng/ml and 0.12 ng/ml was yielded, respectively [72].…”

Advanced Signal-Amplification Strategies for Paper-Based Analytical Devices: A Comprehensive Review

“…Therefore, PEC-based signal amplification enhancement that can address these issues is necessary in order to minimize the number of assays, but still reach high sensitivity with a very low concentration. In the last few years, diverse strategies have been proposed to amplify photoactive signals for PEC-sensing fabrication, most of which have been employed in electron acceptor/donor regulation [ 120 , 123 , 124 , 125 , 126 ], photoactive materials, nanozymes [ 119 , 120 , 127 , 128 , 129 , 130 , 131 , 132 , 133 , 134 , 135 ], or light sources [ 136 , 137 , 138 , 139 , 140 , 141 , 142 ]. Among them, functional nanomaterial integration is an emerging potent route to reach giant, efficient lab-on-paper analytical devices, and diverse photoactive nanomaterials have been immobilized on cellulose fibers.…”

“…Although it is well known that an excited light source is requisite for a photoelectrochemical platform, physical light source-related instruments are also required [ 142 ]. Therefore, to achieve instrument miniaturization as well as operational simplification, luminol-based chemiluminescence emission was exploited as an interior light source, which offers multitudinous advantages, including low cost, simplicity, high sensitivity, and a rapid response [ 136 ]. Lan et al designed and constructed a novel 3D-rGO/cellulose-based photoelectrical lab-on-paper analytical device to determine thrombin (TRB) in serum samples, a procedure typically utilized for Alzheimer disease and cardiovascular disease diagnosis [ 142 ].…”

Recent Advances in Microfluidic Paper-Based Analytical Devices toward High-Throughput Screening

“…For detection, this technique can reduce background noise without the need for an external light source, but a portable chemiluminescence reader must be operated in the dark, which makes device fabrication more complex. This detection method was used with μPADs for biomarker detection [ 145 ], immunoassay [ 146 ], biomolecule detection [ 147 ], environmental monitoring [ 148 ], and food safety control [ 149 ]. In recent years, chemiluminescence sensing has become popular for clinical biomarker detection and immunoassays due to its wide dynamic range and complete automation.…”

“…Yang et al demonstrated a novel paper-based chemiluminescence analytical device based on immunocomplex formation for the simultaneous detection of heart disease biomarkers. This device could be applied for simple, sensitive, and selective detection of three biomarkers in human serum and had low detection limits in the picogram range [ 145 ]. Guo et al fabricated a μPAD by using a dipping method based on a chemiluminescence immunoreaction system for multiplexed detection of three cancer biomarkers in human serum samples [ 149 ].…”