Master of Science
Dr Ken Yeung
To analyze the proteins present in biological samples several major steps are involved. One of the crucial steps in proteomics has been sample preparation. Prior to mass spectral analysis proteins containing disulfide bonds must be reduced which is followed by digestion. In this work, a novel way of performing protein reduction and digestion at nano-liter volume using capillary electrophoresis is presented. In this work, zone passing CE technique was utilized to analyze the reduction products in CE. After the successful in-capillary reduction, digestion of protein was also performed. A pH mediated stacking was used to bring the reduced protein and trypsin together for digestion. For performing in-capillary reactions, very low amount of sample is needed resulting in the miniaturization of the reaction. MALDI MS was used for detection considering the ease of coupling to the instrument and only nano-liter sample volume was needed for MS analysis.
Nazeema, Thasneen Naina, "In-Capillary Reduction and Digestion of Proteins at Nano-Liter Volume Using Capillary Electrophoresis" (2012). University of Western Ontario - Electronic Thesis and Dissertation Repository. Paper 892.