Initially chromatographic techniques were used to separate substances based on their color as was the case with herbal pigments. With time its application area was extended considerably. Nowadays, chromatography is accepted as an extremely sensitive, and effective separation method. Column chromatography is one of the useful separation, and determination methods.
Column chromatography is a protein purification method realized especially based on one of the characteristic features of proteins. Besides, these methods are used to control purity of a protein. HPLC technique which has many superior features including especially its higher sensitivity, rapid turnover rate, its use as a quantitative method, can purify amino acids, proteins, nucleic acids, hydrocarbons, carbohydrates, drugs, antibiotics, and steroids.
Conflict of Interest: None declared. Financial Disclosure: The authors declared that this study has received no financial support. National Center for Biotechnology Information , U. Journal List North Clin Istanb v. North Clin Istanb. Published online Nov Ozlem Coskun.
Author information Article notes Copyright and License information Disclaimer. Correspondence: Dr. Received Feb 17; Accepted Oct 1. This article has been cited by other articles in PMC. Abstract Chromatography is an important biophysical technique that enables the separation, identification, and purification of the components of a mixture for qualitative and quantitative analysis. Keywords: Chromatography , column chromatography , protein purification. Types of chromatography Column chromatography Ion-exchange chromatography Gel-permeation molecular sieve chromatography Affinity chromatography Paper chromatography Thin-layer chromatography Gas chromatography Dye-ligand chromatography Hydrophobic interaction chromatography Pseudoaffinity chromatography High-pressure liquid chromatography HPLC.
Column chromatography Since proteins have difference characteristic features as size, shape, net charge, stationary phase used, and binding capacity, each one of these characteristic components can be purified using chromatographic methods. Open in a separate window. Ion- exchange chromatography Ion- exchange chromatography is based on electrostatic interactions between charged protein groups, and solid support material matrix.
Gel- permeation molecular sieve chromatography The basic principle of this method is to use dextran containing materials to separate macromolecules based on their differences in molecular sizes. Affinity chromatography This chromatography technique is used for the purification of enzymes, hormones, antibodies, nucleic acids, and specific proteins [ 13 ].
Paper chromatography In paper chromatography support material consists of a layer of cellulose highly saturated with water. Gas chromatography In this method stationary phase is a column which is placed in the device, and contains a liquid stationary phase which is adsorbed onto the surface of an inert solid.
Dye- ligand chromatography Development of this technique was based on the demonstration of the ability of many enzymes to bind purine nucleotides for Cibacron Blue F3GA dye [ 19 ]. Hydrophobic interaction chromatography HIC In this method the adsorbents prepared as column material for the ligand binding in affinity chromatography are used. Pseudoaffinity chromatography Some compounds as anthraquinone dyes, and azo-dyes can be used as ligands because of their affinity especially for dehydrogenases, kinases, transferases, and reductases The mostly known type of this kind of chromatography is immobilized metal affinity chromatography IMAC [ 24 ].
Application areas of chromatography in medicine Chromatography technique is a valuable tool for biochemists, besides it can be applied easily during studies performed in clinical laboratories For instance, paper chromatography is used to determine some types of sugar, and amino acids in bodily fluids which are associated with hereditary metabolic disorders.
Conclusion Initially chromatographic techniques were used to separate substances based on their color as was the case with herbal pigments. Footnotes Conflict of Interest: None declared. Selective enzyme purification by affinity chromatography. Porath J. From gel filtration to adsorptive size exclusion. J Protein Chem. Harris DC. Exploring chemical analysis. Preparative ion-exchange chromatography of proteins from dairy whey.
J Chromatogr A. Introduction to organic laboratory techniques. Experimental organic chemistry:Principles and Practice. Oxford:Blacwell Science; —5. Das M, Dasgupta D. Pseudo-affinity column chromatography based rapid purification procedure for T7 RNA polymerase. Prep Biochem Biotechnol. Principles, High Resolution Methods, and Applications. Ion exchange chromatography. New York: Wiley; Amercham Biosciences. Ion Exchange chromatohgraphy, Principles and methods, Amercham Pharmacia.
Biotech SE. Walls D, Loughran ST. Protein chromatography:Methods and protocols, methods in molecular biology. Helmut D. Gel Chromatography, gel filtration, gel permeation, molecular sieves:a laboratory hand book. Springer-Verlag; Determann H. Gel chromatography gel filtration, gel permeation, molecular sieves:a laboratory handbook.
Chapter 2. Materials and Methods. Wilchek M, Chaiken I. An overview of affinity chromatography in affinity chromatography—Methods and protocols. Humana Press. Firer MA. Efficient elution of functional proteins in affinity chromatography. J Biochem Biophys Methods.
TLC plates as a convenient platform for solvent-free reactions. Chem Commun Camb ; 12 —1. Engel introduction to organic laboratory techniques. Amicon, Dye-ligand chromatography. Applications method.
Theory of matrix gel media, Amicon Division. Scopes RK. Use of differential dye-ligand chromatography with affinity elution for enzyme purificationketodeoxyphosphogluconate aldolase from Zymomonas mobilis. Anal Biochem. Cutler P. Methods in molecular biology, Dye-ligand affinity chromatography. Totowa, NJ: Humana Press; Mahn A, Asenjo JA. Prediction of protein retention in hydrophobic interaction Chromatography.
Biotechnol Adv. Hydrophobic interaction chromatography of proteins. J Biotechnol. In a stationary phase, at distinct points the distinct components of a compound are absorbed and also the distinct components prevent moving with mobile phase. This is the procedure by which the results of chromatography are achieved. The mobile phase acts as a solvent in the paper and thin-layer chromatography. A piece of paper is placed in a solvent and that acts as a stationary phase in paper chromatography.
A thin-layer cell plays the role of stationary phase in thin-layer chromatography. These kinds of chromatography avail capillary action to travel the solvent via the stationary phase. The diagrammatic representation of retention factor is as explained below:. Fig1: Explaining Retention Factor. The distinct types of chromatography are as explained below:. The below table explains the chromatography and its applications in the real world and it is as follows:.
Fig2: Table about Chromatography and its Applications. Also See: Hypothesis ppt. All you need to do is just click on the download link and get it. Chromatography PPT Download. If you liked it then please share it or if you want to ask anything then please hit the comment button.
Your email address will not be published. This site uses Akismet to reduce spam. Learn how your comment data is processed. Related Posts. I want ppt for distillation and liquid liquid extraction Reply. Leave a Reply Cancel reply Your email address will not be published. It is used in testing the water samples to know the pollution.
This type of chromatography is availed in the forensics lab to compare the fibers which are found on a victim body, detect bombs in airports and also used in identifying and quantifying the drugs like alcohol.
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