Expression design and optimization of full-length monoclonal antibodies in Pichia pastoris
10.13200/j.cnki.cjb.004462
- VernacularTitle:毕赤酵母表达全长单克隆抗体的设计及优化
- Author:
YE Kaixiong
- Publication Type:Journal Article
- From:
Chinese Journal of Biologicals
2025;38(04):427-435
- CountryChina
- Language:Chinese
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Abstract:
Objective To express monoclonal antibodies(mAbs) in Pichia pastoris chassis host and explore various expression optimization strategies to improve the yield of antibodies.Methods Pichia pastoriswas utilized as the chassis cell for the recombinant expression of various mAbs using both methanol-induced system PAOX1 and ethanol-induced system E1.Molecular chaperone overexpression and media component optimization were employed to further enhance the production of mAbs. The productivity of the recombinant strains was then evaluated at a 3 L reactor scale.Results Ethanol-induced expression of Eptinezumab resulted in a yield of 12. 4 mg/L. Overexpression of molecular chaperones Ppi1 and Biph increased Eptinezumab production by approximately 22. 5% and 23. 0%, respectively. Additionally, the supplementation of1. 0 g/L ammonium sulfate, the adjustment of 0. 05 mol/L potassium phosphate buffer, and the addition of 80 mmol/L threonine individually enhanced Eptinezumab yields by approximately 57. 1%, 25. 8%, and 58. 1%, respectively. At the 3 L bioreactor scale, the Biph-overexpressing strain exhibited a remarkable increase in complete antibody production, achieving a maximum yield of 74. 1 mg/L, which was approximately 5. 9 times higher than that obtained at the flask level.Conclusion Eptinezumab can be successfully expressed in the Pichia pastoris chassis host, and the application of various expression optimization strategies has effectively enhanced the production of the antibody. This study provides novel insights and directions for the expression of monoclonal antibodies in Pichia pastoris.
