Method for target protein identification using thermal stability shift-based fluorescence difference in two-dimensional gel electrophoresis

A method for identifying a target protein of a drug molecule, comprising the steps of:

• (a) preparing mixture A comprising cell lysates or human-derived cells;
• (b) preparing mixture B comprising the mixture of cell lysates or human-derived cells, and a drug molecule;
• (c) modifying the temperature of mixture A and mixture B to a specific and same temperature;
• (d) mixing each of mixture A and mixture B at a specific temperature as obtained in step (c) with fluorescent materials having a different wavelength from each other, respectively, so that the proteins existed in a soluble fraction of each of mixture A and mixture B are labelled with the fluorescent materials having a different wavelength from each other, respectively;
• (e) mixing mixture A and mixture B as obtained in step (d) to prepare mixture C;
• (f) performing electrophoresis of mixture C;
• and
• (g) analyzing a fluorescence wavelength of a protein spot shown in a gel by the electrophoresis of step (f) to confirm a protein showing thermal stability shift resulting from step (c).