Large-scale production of active recombinant human C1q/TNFα related protein-2 in Pichia pastoris

Aim To achieve high-level expression of recombinant hCTRP2 in Pichia pastoris via high-density fermentation.Methods Recombinant hCTRP2 was expressed in Pichia pastoris by high-density fermentation,purified by Ni-NTA affinity chromatography and the bioactivities of purified rhCTRP2 were assayed in vitro test and in vivo test.Results Recombinant transformants with high-level expression of rhCTRP2 were identified and 560 mg·L-1total protein was secreted in 14-L fermentor induction with methanol.The recombinant hCTRP2 proteins were purified using NiNTA affinity chromatography with a yield of about 10.4 mg purified protein from 100 ml culture supernatants.The purified recombinant was assayed to be active by in vitro test and in vivo test.Conclusion The active recombinant hCTRP2 is high-level expressed by fedbatch fermentation and purified with Ni-NTA affinity chromatography.