Case Studies - Bacterial Expression
1. Protein refolding
Overproduction of proteins in E.coli can cause the formation of inactive, misfolded and insoluble protein aggregates. GenScript provides proprietary refolding technology to solve this problem. A case study shows that over 95% of the inclusion body is solublized and refolded. The purity of the refolded protein is more than 85%. The refolded protein is delivered in customized buffer.
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Fig. Refolded protein of sample was analyzed by SDS-PAGE followed by Coomassie Brilliant Blue staining
Lane M: Smart Protein Standard (GenScript, Cat. No. MM0900)
Lane 1: Refolded protein of sample |
2. Tag removal
Presence of epitope tag in recombinant proteins may result in changes in protein structure, toxicity, and loss of bioactivity. To avoid these problems, tag removal after protein expression is one of additional requests from some customers. Shown below is protein that is cleaved by SUMO protease and successfully isolated. About 80% tag free target protein with purity of >90% is recovered.
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SDS-PAGE checking purification of protein after tag cleavage by SUMO protease
Fig. A:
Lane M: Smart Protein Standard (GenScript, Cat.No. MM0900)
Lane 1: Refolded fusion protein with SUMO-tag
Fig. B:
Lane M: Smart Protein Standard (GenScript, Cat.No. MM0900)
Lane 1: Recovered protein after 1st incubation with Ni-resin
Lane 2: Recovered protein after 2nd incubation with Ni-resin
Lane 3: Elute with 300 mM imidazole from Ni-resin of 2nd incubation |
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