| CPC C07K 1/16 (2013.01) [C07K 16/065 (2013.01); C07K 16/4291 (2013.01)] | 29 Claims |

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1. A process of purifying a protein of interest from a protein mixture comprising the protein of interest and a low molecular weight (LMW) variant 2H1L or HH, the process comprising:
a) loading the protein mixture onto Ceramic Hydroxy apatite (CHT) column with a loading buffer;
b) washing the CHT column with a washing buffer; and
c) eluting by a linear gradient with an eluting buffer, thereby producing an eluted protein mixture comprising a purified protein of interest;
wherein the washing buffer is sodium phosphate buffer and the eluting buffer is sodium phosphate buffer;
wherein the eluting buffer comprises sodium phosphate concentration from about 24 mM to about 104 mM; and
wherein the purified protein of interest is substantially pure and 2H1L is less than 3% measured by CE-SDS or HH is less than 0.2% measured by CE-SDS and wherein the purified protein of interest is substantially pure.
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