Abstract：
:In order to develop a serum-free (SF) medium for the production of erythropoietin (EPO) by suspension culture of recombinant Chinese hamster ovary (rCHO) cells, a statistical optimization approach based on a Plackett-Burman design was adopted. A basal medium was prepared by supplementing Iscove's modified Dulbecco's medium (IMDM) with Fe(NO3)3.9H2O, CuCl2 and ZnSO4.7H2O which are generally contained in SF medium formulations. Insulin, transferrin and ethanolamine were also supplemented to the basal medium to determine their optimal concentrations. From this statistical analysis, glutamate, serine, methionine, phosphatidylcholine, hydrocortisone and pluronic F68 were identified as positive determinants for cell growth. The SF medium was formulated by supplementing the basal medium with components showing positive effects on cell growth in suspension culture. An EPO titer in this optimized SF medium was 79% of that in IMDM supplemented with 5% dialyzed fetal bovine serum (dFBS). Furthermore, the in vitro and in vivo biological activities of EPO produced in the SF medium were comparable to those produced in the serum-supplemented medium. Taken together, the results obtained here show that a Plackett-Burman design facilitates the development of SF media for the production of EPO by suspension culture of rCHO cells.
journal_name
J Biotechnoljournal_title
Journal of biotechnologyauthors
Lee GM,Kim EJ,Kim NS,Yoon SK,Ahn YH,Song JYdoi
10.1016/s0168-1656(99)00004-8subject
Has Abstractpub_date
1999-04-15 00:00:00pages
85-93issue
2-3eissn
0168-1656issn
1873-4863pii
S0168-1656(99)00004-8journal_volume
69pub_type
杂志文章abstract：:Combining the isochorismate synthase EntC and the chorismatase FkbO in a sequential enzyme cascade provides a useful system for the biocatalytic production and subsequent purification of isochorismate from an isochorismate/chorismate mixture. FkbO has a strict preference for chorismate - isochorismate is not accepted ...
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pub_type： 杂志文章,评审
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journal_title：Journal of biotechnology
pub_type： 杂志文章,评审
doi：10.1016/0168-1656(92)90114-o
更新日期：1992-08-01 00:00:00
