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Lecture

Protein Mass Spectrometry and Proteomics

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.

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Instructor

Loïc Dayon

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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.

Gas chromatography

Gas chromatography (GC) is a common type of chromatography used in analytical chemistry for separating and analyzing compounds that can be vaporized without decomposition. Typical uses of GC include testing the purity of a particular substance, or separating the different components of a mixture. In preparative chromatography, GC can be used to prepare pure compounds from a mixture. Gas chromatography is also sometimes known as vapor-phase chromatography (VPC), or gas–liquid partition chromatography (GLPC).

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.

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.

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