Provided herein are methods of reducing high molecular weight (HMW) species and/or high mannose species in a protein sample. The present invention provides a purification method for removal of high molecular weight species from a protein sample using two sequential chromatographic steps. When developing processes for commercial production of therapeutically important proteins, purification of the protein sample to remove undesired species in an efficient manner is highly desirable. Purification processes include procedures in which the protein of interest is separated from the source materials on the basis of solubility, ionic charge, molecular size, adsorption properties, and specific binding to other molecules.
A number of well-known technologies are utilized for purifying desired proteins from their source material. The recombinant proteins are then either secreted into the surrounding media or into the perinuclear space, or made intracellularly and extracted from the cells. Therapeutic proteins may be obtained from plasma or tissue extracts, for example, or may be produced by recombinant methods such as by cultures of eukaryotic or prokaryotic cells containing at least one recombinant plasmid encoding the desired protein.
The purification of proteins for the production of biological or pharmaceutical products from various source materials involves a number of procedures. The present disclosure is directed to removal or decrease in amount of high molecular weight species and/or high mannose species from a protein sample using a purification comprising a mixed mode chromatography step and a hydrophobic interaction chromatography step. 13, 2013 is claimed, the disclosure of which is incorporated by reference in its entirety.
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