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Imunochemická detekce proteinu TNF-α pomocí magnetických částic
Koudelková, Žaneta
This thesis is focused on development and optimization of a method for detection of the cytokine TNF-α by the sandwich ELISA using magnetic particles. These particles were functionalized with carboxylic functional groups to enable the antibody binding. Optimal pH of the MES buffer, EDC concentration, and method of activation were determined by optimizing the conditions of particle activation. In the second part of the thesis, the conditions of the magnetic particle-based sandwich ELISA were optimized. It was discovered that mixing the samples during incubation resulted in a higher number of molecules being captured by the antibodies. The optimal incubation times for the antigen and detection antibody were determined to be 3 hours and 1.5 hours, respectively. The optimized conditions for particle activation and ELISA were used to create a calibration curve of TNF-α standards. Optimized magnetic particle-based ELISA protocol can simplify and enhance the analysis of complex samples like blood serum, urine, or saliva.

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