KIN217 Lecture Notes - Lecture 3: Affinity Chromatography, Differential Centrifugation, Protein Purification
23 Sep 2018
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Kin 217 lecture 3 protein analysis and enzymes. Proteome functional presentation of the genome; the study of proteins. Usually involves protein purification and mass spectrometry analyses. More complex than genomics: genome fixed; represent a list of gene products, proteome not fixed; represents the functional expression of information, and varies with cell type, developmental stage and environment condition. Proteins are purified on the basis of differences in solubility, size, charge, and specific binding affinity. Cells are disrupted in a homogenizer, and the resulting mixture (homogenate) is centrifuged in a step-by-step fashion of increasing centrifugal force. The denser material will form a pellet at lower centrifugal force than the less dense material. The isolated fractions can be used for further purification. Salting out: proteins precipitate out of solution at high salt concentrations, collect precipitate and then remove salt (dialysis) Separation by size: molecular exclusion chromatography, aka gel filtration chromatography. Small molecules enter porous beads and take a longer path, will exit last.
