PHAR 505 Lecture Notes - Lecture 4: T7 Rna Polymerase, Lac Repressor, Multiple Cloning Site
23 Nov 2017
School
Department
Course
Professor
Document Summary
Phar 505 lecture 4 (09/14/2017) by: r. akong. Protein production: e. coli bacteria, eukaryotic systems. Protein purification methods: extraction, principles of chromatography. Other biophysical methods to determine the affinity between a drug and protein also require pure material: this confirms that the drug is effectively hitting the right target and concentrations can be determined accurately for proper kd estimation. Purification enables a better functional characterisation of the protein: more pure = higher specific activity, this is the level of activity per unit mass of the protein. This is essentially biochemistry but a good pharmacologist should know how proteins are prepared and purified. Proteins can be isolated from natural sources: myoglobin from sperm whale muscles. Insulin from animal pancreas: ribosomes from bacteria. Protein needs to be abundant, otherwise large amounts of starting material are required. Proteins can be overexpressed in genetically modified systems such as: e. coli bacteria, baculovirus insect cells, pichia pastoris yeast, mammalian cells hek293, cho cells.
