• Dr. Sakinala padmavathi
• Dr. Shaik Abdul Rahaman
• V. Srividya
• A. Jyothika
• Dr. Anjana male

The diversity in biomolecules makes them difficult to separate in their pure form. In order to augment the biological activity of many important biomolecules it is very necessary to separate and purify them in their purest form. This review deliberates the chromatographic purification of biomolecules as one of the most emerging technology. In this the basic principle for the separation of biomolecules along with different types of chromatography, their principle and applications is discussed. This review also focused on use of combination of two or more chromatographic techniques for the separation of therapeutic proteins like monoclonal antibodies (mAbs) and enzymes to achieve their native activity. Affinity chromatography is a separation method based on specific binding interaction between an immobilised ligand and its binding partner. For example include antibody/antigen, enzyme/ substrate, and enzyme/ inhibitor interaction. Binding to the solid phase may be achieved by column chromatography whereby the solid medium is packed onto a column, the initial mixture run through the column to allow settling, a wash buffer run through the column and the elution buffer subsequently applied to the column and collected. These steps are usually done at ambient pressure. Alternatively, binding may be achieved using a batch treatment, for example, by adding the initial mixture to the solid phase in a vessel, mixing, separating the solid phase, removing the liquid phase, washing, re-centrifuging, adding the elution buffer, re-centrifuging and removing the elute. Sometimes a hybrid method is employed such that the binding is done by the batch method, but the solid phase with the target molecule bound is packed onto a column and washing and elution are done on the column.

Chromatography introduction, column chromatography, Affinity Chromatography, Separation of proteins

"SEPERATION OF PROTEINS BY CHROMATOGRAPHIC TECHNIQUES", International Journal of Emerging Technologies and Innovative Research (www.jetir.org), ISSN:2349-5162, Vol.10, Issue 9, page no.f204-f211, September-2023, Available :http://www.jetir.org/papers/JETIR2309525.pdf

"SEPERATION OF PROTEINS BY CHROMATOGRAPHIC TECHNIQUES", International Journal of Emerging Technologies and Innovative Research (www.jetir.org | UGC and issn Approved), ISSN:2349-5162, Vol.10, Issue 9, page no. ppf204-f211, September-2023, Available at : http://www.jetir.org/papers/JETIR2309525.pdf