Polyelectrolyte Multilayer Substrate Flow Elisa System Research

Enzyme-linked immunosorbent assay (ELISA) is suitable for small amount of protein detection and quantitative test. ELISA is widely used in food, industry, environmental, clinical medicine, and so on. With the advent of various protein markers, the requirements of disease detection are difficult to need. Traditional ELISA has some shortcomings, such as low sensitivity, long incubation time and mixed sample detection difficulty which limited its devolopment. Therefore, in order to realize "rapid-sensitive-simple" immunosorbent detection technology, we developed electrophoresis ELISA system:1. We applied layer-by-layer (LbL) assembly technology on polyelectrolyte multilayers (PEMs) preparation. We reviewed the content of polyelectrolyte composition on PEMs and the stability of PEMs; We also researched the quality of primary antibody and blocking agent adsorption on PEMs.2. In ELISA electrophoresis system, we detected the model antigen (mouse IgG). The limit of detection (LOD) of mouse IgG was 33 pM in optimized electrophoresis condition (25 V,2 min), the detection time was 1/30 shortener than the traditional ELISA.3. C-reactive protein (CRP) as a biomarker of inflammation verified the clinical application of electrophoresis ELISA system. CRP could be separated from complex mixture solution in temporal space by electrophoresis force. The LOD of CRP was 33 pM by electrophoresis detection system.The incubation time for antigen-antibody reaction was just 2 min.4. We combined antigen electrophoresis technology with nano-microsphere electrophoresis technology to realize signal amplification. Finally, the LOD of mouse IgG was 0.13 pM. The sensitivity was improved 254 times than antigen electrophoresis and the reaction time of antigen with primary antibody and antigen with secondary antibody were 1/30 shortener than the traditional ELISA.