Design And Research Of Novel Electrochemical Immunosensor For Lymphoma Biomarkers Based On Synergy Signal Amplification Of Alloy Particles

Objective C-myc and Bcl-2 protein,the biomarker of double expresseor lymphoma,were selected as the target in this paper.Taking the disadvantages of complex detection,low sensitivity,and mostly single target analysis faced by electrochemical immunosensor for lymphoma biomarkers,it is planned to start with identification element fixation substrate,signal markers,and the cooperative signal amplification strategies for the above,in order to establish a series of single and combined lymphoma biomarkers electrochemical immunosensing method with high sensitivity,stability and selectivity.The above research not only provides an efficient and precise methodological basis for trace molecular recognition and molecular diagnosis in vivo,but also effectively improves the sensitivity and detection rate of lymphoma diagnosis.The high-efficiency analysis provides new ideas for rapid screening and testing of population samples.Methods In this paper,a series of co-loaded alloys were prepared by adsorption method and mediator reduction method.Field emission scanning electron microscopy(FESEM),transmission electron microscopy(TEM),dispersive X-ray spectroscopy(EDS),element mapping,fourier infrared spectroscopy(FT-IR)for microscopic morphology characterization,element and active group analysis;differential pulse voltammetry method(DPV)was used to study the electrochemical performance.Four co-loading alloys with excellent electrical behavior,PB-Au Pd,TB-Au Pd,MB-Pt Pd and TMB-Pt Pd were selected and used as detection antibody labels.The electrodeposited electrode(E-Au Pd/GCE),carbon-based alloy particle modified electrode(CS-GO-Au Pd/GCE)and electropolymerized electrodes(PEDOT-GOMWCNTs/GCE)were used as sensing interface to immobilize the capture antibody.The single biomarker C-myc protein and the combined biomarker C-myc and the Bcl-2 protein of the lymphoma,which were identified and detected based on the specific binding by antigen-antibody,respectively.The morphology of the modified electrodes were characterized by FESEM,cyclic voltammetry(CV)and electrochemical impedance spectroscopy(EIS)were used to monitor the process of sensor construction.Differential pulse voltammetry(DPV)was used to detect C-myc and Bcl-2 protein sensitively.The analytical performances of three sensors are systematically studied through methodological research and feasibility validation.Results Quantitative detection ranges of the two electrochemical immunosensors for Cmyc protein were 1 pg/m L～1 ng/m L and 0.5 pg/m L～1 ng/m L,with detection limits of 0.5pg/m L and 0.25 pg/m L,respectively.The linear ranges of simultaneous detection of C-myc and Bcl-2 protein in electrochemical immunosensors are 1 pg/m L～1 ng/m L and 5 pg/m L～1ng/m L,and the detection limits are 1 pg/m L and 2.5 pg/m L,respectively.Systematic methodological validation and feasibility analysis confirmed that the three immunosensors had good stability,reproducibility and selectivity.Three immunosensors were applied to the detection of standard-added blood samples,respectively.The recoveries ranged from 95.3%to 107.7% with RSD less than 4.10%.Conclusions In this paper,alloy particles were applied both in electrode modification and signal markers simultaneously.Two novel electrochemical immunosensors based on synergistic signal amplification technology for the detection of C-myc protein have shown comparable or even better analytical performances than those reported.Then the mediator reduction method is preferably used to prepare different co-loading alloy particles for labeling detection antibody in order to generate distinguishable signals.Also,the alloys were further used for enzyme-free catalysis to amplify the detection signals.Finally,C-myc and Bcl-2 proteins can be detected simultaneously using electrochemical immunosensor.The above immunosensors have satisfactory recovery rate in various standard serum samples,which confirmed that the constructed series of electrochemical immunosensors have potential for application in practical detection.Moreover,the simultaneous analysis of the multiplex biomarkers is realized while ensuring the sensitivity of immunosensor,which can be used as an auxiliary tool to improve the specificity of diagnosis.