Lecture

Protein Mass Spectrometry and Proteomics

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In course

CH-419: Protein mass spectrometry and proteomics

In systems biology, proteomics represents an essential pillar. The understanding of protein function and regulation provides key information to decipher the complexity of living systems. Proteomic technology now enables deep quantitative proteome mapping v ...

Description

This lecture covers the fundamentals of protein mass spectrometry and proteomics, starting with an introduction to chromatography techniques and the history of chromatography. It delves into the basis of chromatography, efficiency, resolution, and factors affecting chromatographic performance. The lecture also discusses the importance of selectivity, efficiency, and resolution in chromatography, as well as the concept of theoretical plates. Additionally, it explores the application of peptide isoelectric focusing (IEF) in mass spectrometry data validation and filtering, including the use of immobilized pH gradient (IPG) strips. The lecture concludes with a discussion on the separation efficiency, visualization, and practical applications of off-gel electrophoresis in proteomics.

Instructor

Loïc Dayon

Teaching & PhD Courses Protein mass spectrometry and proteomics CH-419 In systems biology, proteomics represents an essential pillar. The understanding of protein function and regulation provides key information to decipher the complexity of living systems. Proteomic technology now enables deep quantitative proteome mapping via mass spectrometry.

Official source

Related concepts (32)

Chromatography

In chemical analysis, chromatography is a laboratory technique for the separation of a mixture into its components. The mixture is dissolved in a fluid solvent (gas or liquid) called the mobile phase, which carries it through a system (a column, a capillary tube, a plate, or a sheet) on which a material called the stationary phase is fixed. Because the different constituents of the mixture tend to have different affinities for the stationary phase and are retained for different lengths of time depending on their interactions with its surface sites, the constituents travel at different apparent velocities in the mobile fluid, causing them to separate.

High-performance liquid chromatography

High-performance liquid chromatography (HPLC), formerly referred to as high-pressure liquid chromatography, is a technique in analytical chemistry used to separate, identify, and quantify each component in a mixture. It relies on pumps to pass a pressurized liquid solvent containing the sample mixture through a column filled with a solid adsorbent material. Each component in the sample interacts slightly differently with the adsorbent material, causing different flow rates for the different components and leading to the separation of the components as they flow out of the column.

Paper chromatography

Paper chromatography is an analytical method used to separate coloured chemicals or substances. It is now primarily used as a teaching tool, having been replaced in the laboratory by other chromatography methods such as thin-layer chromatography (TLC). The setup has three components. The mobile phase is a solution that travels up the stationary phase, due to capillary action. The mobile phase is generally a mixture of non-polar organic solvent, while the stationary phase is polar inorganic solvent water.

Mass spectrometry

Mass spectrometry (MS) is an analytical technique that is used to measure the mass-to-charge ratio of ions. The results are presented as a mass spectrum, a plot of intensity as a function of the mass-to-charge ratio. Mass spectrometry is used in many different fields and is applied to pure samples as well as complex mixtures. A mass spectrum is a type of plot of the ion signal as a function of the mass-to-charge ratio.

Column chromatography

Column chromatography in chemistry is a chromatography method used to isolate a single chemical compound from a mixture. Chromatography is able to separate substances based on differential adsorption of compounds to the adsorbent; compounds move through the column at different rates, allowing them to be separated into fractions. The technique is widely applicable, as many different adsorbents (normal phase, reversed phase, or otherwise) can be used with a wide range of solvents.

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Related concepts (32)

Chromatography

High-performance liquid chromatography

Paper chromatography

Mass spectrometry

Column chromatography