Chromatography is used to separate and purify biomolecules according to their difference in specific properties. This method includes a stationary phase to be purified across a mobile phase. This technique is used for the recovery, analysis, purification, and separation of the products. column chromatography is one of the most common methods of protein purification. Chromatography is based on the principle where molecules in mixture applied onto the surface or into the solid, and fluid stationary phase (stable phase) is separating from each other while moving with the aid of a mobile phase