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PROTEIN CHROMATOGRAPHY

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PROTEIN CHROMATOGRAPHY ( protein-chromatography )

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Lavanya G et al / J Biomed Sci and Res., Vol 3 (3), 2011,424-438 PROTEIN CHROMATOGRAPHY *Lavanya.G, Mubarak.SK, Vishali , Sujana.K, Pramela Rani, B. Syama Sundar ANU college of Pharmaceutical sciences, Acharya Nagarjuna University, Guntur dist, Andhra Pradesh, India Abstract: Proteins, peptides, lipids, amino acids, carbohydrates, vitamins and drugs can be separated using chromatography . Chromatography is the separation of a mixture in to individual components using a stationary phase and mobile phase. Proteins can be separated by using chromatographic techniques like size exclusion chromatography, ion exchange chromatography, affinity chromatography, fast protein liquid chromatography, high performance liquid chromatography, reversed phase chromatography, electrophoresis. Separation techniques rely on the differences in the solubility, size, charge, and adsorption characteristics of protein molecules. Ion-exchange chromatography is used to separate proteins on the basis of charge. Affinity chromatography utilises ligands, such as enzyme inhibitors, coenzymes, or antibodies, to specifically bind proteins to a solid support. Size of the Protein can be separated by using size exclusion chromatography. Electrophoresis can be used to separate proteins from complex mixtures on the basis of size and charge. Keywords: affinity chromatography, Chromatography, electrophoresis,fast protein liquid chromatographyhigh performance liquid chromatography, ion exchange chromatography, reversed phase chromatography, size exclusion chromatography. Introduction: Chromatography Chromatography is the separation of a mixture in to individual components using a stationary phase and mobile phase. These include proteins, peptides, lipids, amino acids, carbohydrates, vitamins and drugs. History The credit for the discovery chromatography goes to Russian botanist Mikhail Tswett in 1906. Tswett described the separation of plant leaf pigments in solution by passing through a column of solid adsorbents. He coined the term chromatography (Greek = Chroma-colour; graphin = to write), since the technique dealt with the separation of colour compounds. Principle The principle of separation can be either adsorption or partition. Hence can be called as adsorption chromatography (or) partition chromatography. The stationary phase can be a solid or liquid, while the moving phase is a liquid or gas. When the stationary phase is solid, the basis is adsorption and when it is liquid, the basis is partition. Theory Plate theory and Rate theory are two theories that are applicable to chromatography. 1) Plate Theory: Plate theory describes a chromatography system as being in equilibrium between the stationary and mobile phases. This views the column as divided into a number of imaginary theoretical plate. This is significant because as the number of plates in a column increases (or) HETP increases, so does the separation of components it also provides as equation that describes the elution curve (or) the chromatogram of a solute it can also be used to find the volume and the column efficiency. HETP=L/N Where : L = Column length N = Number of theoretical plates 2) Rate Theory: The Rate theory describes the migration of molecules in a column. This included band shape, broadening and the diffusion of a solute. of 424

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