Case Studies

Case Study: An Example of Protein Expression and Purification from Yeast In our paper on A2E macular degeneration (Moody, et al., 2016) we synthesized recombinant manganese peroxidase in a P. pastoris strain with an integrated rMnP expression construct isolated from P. chrysoporium. The isolated sequence was cloned into a pGAPZa-A expression vector with an n-terminal alpha factor secretion sequence and a c-terminal myc and histidine tag. The protein was expressed in a 2.8L starter culture and transferred to a 14L Eppendorf bioreactor. The protein was purified in a single step nickel-nitrilotriacetic acid (NiNTA) affinity chromatography column. Production of recombinant manganese peroxidase is a representative example of applied techniques we utilize at Ichor Discovery, allowed us to test rMnP efficacy in breaking down lipofuscin flourophores in the search for treatments of macular degeneration. Read the full article linked below. Recombinant Manganese Peroxidase Reduces A2E Burden in Age-Related and Stargardt’s Macular Degeneration…