title: NaCS-PDMDAAC immobilized cultivation of recombinant Dictyostelium discoideum for soluble human Fas ligand production.
abstract: Dictyostelium discoideum is a promising eukaryotic host for the expression of heterologous proteins requiring post-translational modifications. However, the dilute nature of D. discoideum cell culture limits applications for high value proteins production. D. discoideum cells, entrapped in sodium cellulose sulfate/poly-dimethyl-diallyl-ammonium chloride (NaCS-PDMDAAC) capsules were used for biosynthesis of the heterologous protein, soluble human Fas ligand (hFasL). Semi-continuous cultivations with capsules recycling were carried out in shake flasks. Also, a scaled-up cultivation of immobilized D. discoideum for hFasL production in a customized vitreous airlift bioreactor was conducted. The results show that NaCS-PDMDAAC capsules have desirable biophysical properties including biocompatibility with the D. discoideum cells and good mechanical stability throughout the duration of cultivation. A maximum cell density of 2.02 × 10(7) cells mL(-1) (equivalent to a maximum cell density of 2.22 × 10(8) cells mL(-1) in capsules) and a hFasL concentration of 130.40 μg L(-1) (equivalent to a hFasL concentration of 1434.40 μg L(-1) in capsules) were obtained in shake flask cultivation with capsules recycling. Also, a maximum cell density of 1.72 × 10(7) cells mL(-1) (equivalent to a maximum cell density of 1.89 × 10(8) cells mL(-1) in capsules) and a hFasL concentration of 106.10 μg L(-1) (equivalent to a hFasL concentration of 1167.10 μg L(-1) in capsules) were obtained after ∼170 h cultivation in the airlift bioreactor (with a working volume of 200 mL in a 315 mL bioreactor). As the article presents a premier work in the application of NaCS-PDMDAAC immobilized D. discoideum cells for the production of hFasL, more work is required to further optimize the system to generate higher cell densities and hFasL titers for large-scale applications.
authors.affiliation: Dept. of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, People's Republic of China; Scientific Research Center, Zhejiang Inst. of Medical Devices, Hangzhou, 310009, People's Republic of China.
authors.forename: Chao
authors.initials: C
authors.lastname: Zheng
authors.forename: Xianhai
authors.initials: X
authors.lastname: Zeng
authors.forename: Michael K
authors.initials: MK
authors.lastname: Danquah
authors.forename: Yinghua
authors.initials: Y
authors.lastname: Lu
mesh_terms: D001672:Biocompatible Materials / Q000737:chemistry / Q000633:toxicity; D019149:Bioreactors / Q000469:parasitology*; D018914:Cells, Immobilized / Q000737:chemistry / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D002482:Cellulose / Q000031:analogs & derivatives* / Q000737:chemistry / Q000633:toxicity; D004023:Dictyostelium / Q000166:cytology / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D053222:Fas Ligand Protein / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D011095:Polyethylenes / Q000737:chemistry* / Q000633:toxicity; D000644:Quaternary Ammonium Compounds / Q000737:chemistry* / Q000633:toxicity
chemical_list: D001672:Biocompatible Materials; D053222:Fas Ligand Protein; D011095:Polyethylenes; D000644:Quaternary Ammonium Compounds; C041004:poly-N,N-dimethyl-N,N-diallylammonium chloride; D002482:Cellulose; C020254:cellulose sulfate
keywords: Dictyostelium discoideum; NaCS-PDMDAAC capsules; immobilized cultivation; soluble human Fas ligand