An organic solvent-based protocol for depletion of abundant proteins in plasma of Leprosy patients

Abstract In present study protein was purified from human plasma with the method involving organic solvents. Organic solvents like chloroform, methanol and isopropanol remove the lipoproteins and glycoproteins whereas TCA and Acetone are known to deplete the abundant proteins like albumins and globulins from the plasma of the blood. Purified protein was subjected to the estimation by Bradford Assay (96 well plate method). In this process a dye called Brilliant Blue forms complex with the purified protein of plasma because of which the maximum absorption of dye shifts to 595nm. So, the absorbance was observed at wavelength of 595nm. Purified protein along with unpurfled plasma was imposed on SDS-PAGE. SDS (sodium dodecyl sulphate) is an anionic detergent which first denatures the protein and then provides all the protein fragments a negative charge. After which polyacrylamide gel the separates the protein fragments on the basis of their molecular mass. Results of SDS-PAGE concluded that the protein was purified when its bands were compared to that of unpurfled plasma, which means the treatment with organic solvents is useful in purifying protein from plasma of leprosy patient.