title: The expression and characterization of functionally active soluble CD83 by Pichia pastoris using high-density fermentation.
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.
authors.affiliation: Hefei 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.
authors.forename: Yugang
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Hefei 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.
authors.forename: Rui
authors.initials: R
authors.lastname: Li
authors.affiliation: Department of Pharmacy, Anhui Medical College, Hefei, China.
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Song
authors.affiliation: Hefei 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.
authors.forename: Yongjun
authors.initials: Y
authors.lastname: Zhong
authors.affiliation: Hefei 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.
authors.forename: Chenguang
authors.initials: C
authors.lastname: Wang
authors.affiliation: Hefei 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.
authors.forename: Hao
authors.initials: H
authors.lastname: Jia
authors.affiliation: Hefei 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.
authors.forename: Lidan
authors.initials: L
authors.lastname: Wu
authors.affiliation: Hefei 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.
authors.forename: Dong
authors.initials: D
authors.lastname: Wang
authors.affiliation: Hefei 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.
authors.forename: Fang
authors.initials: F
authors.lastname: Fang
authors.affiliation: Center of Medical Biotechnology of Anhui Province, University of Science and Technology of China, Hefei, China.
authors.forename: Jiajia
authors.initials: J
authors.lastname: Ma
authors.affiliation: Hefei 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.
authors.forename: Wenyao
authors.initials: W
authors.lastname: Kang
authors.affiliation: Hefei 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.
authors.forename: Jie
authors.initials: J
authors.lastname: Sun
authors.affiliation: Hefei 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.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Tian
authors.affiliation: Hefei 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.
authors.forename: Weihua
authors.initials: W
authors.lastname: Xiao
mesh_terms: D015703:Antigens, CD / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D015153:Blotting, Western; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000166:cytology* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005285:Fermentation; D006801:Humans; D007136:Immunoglobulins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D009000:Monocytes / Q000166:cytology* / Q000378:metabolism; D010843:Pichia / Q000254:growth & development / Q000378:metabolism*; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000097802:CD83 Antigen
chemical_list: D015703:Antigens, CD; D007136:Immunoglobulins; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D000097802:CD83 Antigen