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Updated: May 2, 2026
Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris
Published on: February 25, 2010
The expression and characterization of functionally active soluble CD83 by Pichia pastoris using high-density
Yugang Guo1, Rui Li2, Xiaoping Song3
1Hefei National Laboratory for Physical Sciences at Microscale, Innovation Center for Cell Biology, School of Life Sciences, University of Science and Technology of China, Hefei, China ; Center of Medical Biotechnology of Anhui Province, University of Science and Technology of China, Hefei, China.
Insights
Researchers produced soluble CD83 (sCD83) using Pichia pastoris fermentation. This functional immune suppressor effectively inhibited dendritic cell maturation and T-cell proliferation, showing potential for clinical applications.
Area of Science:
- Immunology
- Biochemistry
- Molecular Biology
Background:
- CD83 is a key glycoprotein in dendritic cell (DC) maturation and adaptive immunity initiation.
- The soluble isoform, sCD83, acts as an immune suppressor by inhibiting DC maturation.
- Efficient production of functional sCD83 is crucial for therapeutic development.
Purpose of the Study:
- To develop a high-yield strategy for expressing functional soluble CD83 (sCD83) using Pichia pastoris.
- To characterize the properties and biological activity of recombinant sCD83.
- To evaluate the potential of yeast-expressed sCD83 for immunological studies and clinical applications.
Main Methods:
- High-density fermentation of Pichia pastoris for sCD83 expression.
- Purification and characterization of recombinant sCD83, including glycosylation analysis (PNGase F digestion).
- In vitro assays to assess sCD83 binding to monocytes and blocking of anti-CD83 antibodies on DCs.
- Evaluation of sCD83's effect on ConA-stimulated peripheral blood mononuclear cell (PBMC) proliferation.
Main Results:
- Achieved high-yield (over 200 mg/L) monomeric sCD83 expression in Pichia pastoris.
- Confirmed N-linked glycosylation sites on the purified recombinant sCD83.
- Demonstrated that recombinant sCD83 binds to monocytes and blocks anti-CD83 antibody binding to DCs.
- Showed significant suppression of PBMC proliferation by yeast-expressed sCD83.
Conclusions:
- Pichia pastoris is a suitable host for producing functionally active, soluble CD83 (sCD83).
- Recombinant sCD83 exhibits immunosuppressive properties, validating its potential as a therapeutic agent.
- The developed expression strategy enables further in vivo/in vitro studies and potential clinical translation of sCD83.
Abstract:
CD83 is a highly glycosylated type I transmembrane glycoprotein that belongs to the immunoglobulin superfamily. CD83 is upregulated during dendritic cell (DC) maturation, which is critical for the initiation of adaptive immune responses. The soluble isoform of CD83 (sCD83) is encoded by alternative splicing from full-length CD83 mRNA and inhibits DC maturation, which suggests that sCD83 acts as a potential immune suppressor. In this study, we developed a sound strategy to express functional sCD83 from Pichia pastoris in extremely high-density fermentation. Purified sCD83 was expressed as a monomer at a yield of more than 200 mg/L and contained N-linked glycosylation sites that were characterized by PNGase F digestion. In vitro tests indicated that recombinant sCD83 bound to its putative counterpart on monocytes and specifically blocked the binding of anti-CD83 antibodies to cell surface CD83 on DCs. Moreover, sCD83 from yeast significantly suppressed ConA-stimulated PBMC proliferation. Therefore, sCD83 that was expressed from the P. pastoris was functionally active and may be used for in vivo and in vitro studies as well as future clinical applications.
