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pharmacokinetics anti-angiogenic effects

Materials and Methods

Expression and Purification of Human Irisin

The production of human irisin was performed as previously described [8]. Briefly, human irisin cDNA (360bp) was synthesized by Life Technologies. After cloning the cDNA into EcoR1/Xba1 sites of the pPICZαA plasmid, the linearized pPICZαA-irisin was transformed into Pichia pastoris X-33. The resulting transformant was cultured and induced as previously described [16]. The recombinant irisin was first precipitated via addition of ammonium sulfate and then further purified by loading the precipitated protein onto a concanavalin A–agarose column as introduced by our previous study [8].