Long-term production of soluble human Fas ligand through immobilization of Dictyostelium discoideum in a fibrous bed bioreactor.
The production of recombinant glycoproteins in Dictyostelium discoideum by conventional cell culture methods was limited by low cell density as well as low growth rate. In this work, cotton towel with a good adsorption capability for D. discoideum cells was used as the immobilization matrix in an external fibrous bed bioreactor (FBB) system. With batch cultures in the FBB, the concentration of immobilized cells in the cotton fiber carrier increased to 1.37 x 10(8) cells per milliliter after 110-h cultivation, which was about tenfold higher than the maximal cell density in the conventional free-cell culture. Correspondingly, a high concentration of soluble human Fas ligand (hFasL; 173.7 microg l(-1)) was achieved with a high productivity (23 microg l(-1) h(-1)). The FBB system also maintained a high density of viable cells for hFasL production during repeated-batch cultures, achieving a productivity of 9 approximately 10 microg l(-1) h(-1) in all three batches studied during 15 days. The repeated-batch culture using immobilized cells of D. discoideum in the FBB system thus provides a good method for long-term and high-level production of hFasL.
Chen
Jie
J
Department of Chemical Engineering and Bioengineering, Zhejiang University, Hangzhou 310027, People's Republic of China.
Chen
Heming
H
Zhu
Xiangchen
X
Lu
Yinghua
Y
Yang
Shang-Tian
ST
Xu
Zhinan
Z
Cen
Peilin
P
D000818:Animals; D019149:Bioreactors / Q000382:microbiology*; D001709:Biotechnology; D018914:Cells, Immobilized / Q000378:metabolism*; D004023:Dictyostelium / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D053222:Fas Ligand Protein / Q000235:genetics / Q000378:metabolism*; D005285:Fermentation; D006801:Humans; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012995:Solubility
D053222:Fas Ligand Protein; D011994:Recombinant Proteins