Protein aggregation is a biological phenomenon. There are many reasons why the aggregates form. Mostly the mis-folded proteins have a tendency to form aggregate. Both intra- or extra-cellular protein aggregates are found. These protein aggregates are often toxic; protein aggregates have been implicated in a wide variety of disease known as amyloidosis, including Alzheimer's, Parkinson's and prion disease.Nowadays, monoclonal antibody proteins are widely being used in the field of bio therapeutics. So these bio therapeutic proteins must be free from aggregates. Separation of the pure antibody monomer needs to be very well resolved from its dimer and higher order aggregates. Similarly for quality control and regulatory purpose the separation of antibody fragments is also very much essential. The species other than the monomer might induce toxic side effects to the body if not removed. We have developed TSKgel UltraSW Aggregate column, which is a silica-based 7.8 mm ID � 30 cm analytical SEC column packed with newly developed 30 nm pore size, 3 �m particles. This column is designed especially for mAb analysis particularly for the separation of the aggregates. Larger pore size with the estimated exclusion limit of ~2 x 106Da provides improved separation and quantitation of mAb aggregates and oligomers.We used a number of monoclonal antibodies and proteins of variety of sizes and hydrophobicities to in separating the corresponding protein aggregates. The aggregates are formed by forced denaturation using heat. This study shows that the novel TSKgel UltraSW Aggregate SEC column is suitable for the analysis and purification of antibodies and other proteins from their aggregate and fragment impurities.

Separation of monoclonal antibody and protein aggregates induced by thermal denaturation, using a novel size exclusion chromatography column

246th National Meeting (2013)

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