Analysis of Peptide Quantification and Digestion through Assays, HPLC, and Modification of Workflow

Abstract

The goal of the project is to find a believable and accurate method to quantify peptides, by using a variety of commercial protein assays and HPLC. In the field of proteomics, mass spectrometry is the premier method for attaining the mass to charge ratio of a protein. Proteins must be applied to the mass spectrometer as digested peptide fragments. Quantification of peptide samples is therefore an important topic in both the preparation of samples to apply, as well as important information of unknown samples. Currently there is no method by which peptide concentration can be precisely found. Through the application and modification of already existing protein assays as well as HPLC methods, the sensitivity of the assay and technology for peptide samples will be measured. Methods in the workflow leading up to mass spectrometry must be considered as well such as purification of samples, robustness of methods to solubilize proteins and the methods by with proteins are digested.