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第
19
卷第
6
期
2005
年
12
月
高校化学工程学报
Jom
咀
al
ofChemical
Engineering
ofChinese
Universities
文章编号
1003-9015(2005)06-0855-04
用高效廉价载体培养
CHO
细胞生产
rHuEPO
的研究
陈英文黄任远
1
,
2
沈树宝
高
坤
2
胡云龙
2
(1.南京工业大学
制药与生命科学学院,
江苏南京
210009;
2.
南京华欣药业生物工程有限公司,
江苏南京
210002
)
NO.6
Vo
l.l
9
Dec.
2005
摘
要
z
研究了以自制胶原蛋白载体及
C-DISK
载体培养
CHO
细胞生产
rHuEPO
蛋白的过程,并以进口A-
DISK
载
体作为对照培养。对培养过程中细胞贴壁过程、葡萄糖代谢速率、
rH
uE
PO
浓度及葡萄糖代谢速率与
rH
uE
PO
的浓度
变化关系进行了实验研究分析和对比。结果表明:自制胶原蛋白载体培养细胞具有良好的生物代谢活力和蛋白表达能
力,葡萄糖代谢速率高达
14.2
mmol.
mL→
.h-
1
,比自制
C-DISK
载体高
16
.4%,比进口
A-DISK
载体高
12.7%;
在自制
胶原蛋白载体上培养
CHO
细胞时
EPO
表达量也最高,培养
10
天后
EPO
浓度达到
282
U.
mL-
1
,比自制
C-DISK
载体
高
30%
,也比进口
A-DISK
载体高
18%
。自制胶原蛋白载体具有较佳的性价比,是一种高效、廉价的动物细胞培养载
体。细胞的葡萄糖代谢水平对蛋白表达有着重要的影响,但两者又不同步,因此可用葡萄糖的代谢速率变化对该培养
过程和目的蛋白表达进行适当监测和调控。
关键词
2
细胞培养:载体
CHO
动物细胞
rH
uE
PO(
重组人红细胞生长素)
中图分类号
Q813.11;
Q343.6; TQ934
文献析、识码
A
Production
of
rHuEPO
via
Culturing
CHO
Cells
on
High
Efficient
and
Cheap
Culture
Carrier
CHEN Ying-wen
I,
HUANG
Ren-'
沪Ja
n
口,
SHEN Shu-bao
I,
GAO Kun
2
, HU Yun-long
2
(1.
College
ofPhannacy
and
Li
fe
Science, N
叫
ing
University ofTechnology, Nanjing 210009, China;
2. Nanjing Huaxin Bio-pharmaceutical
Lt
d.
, 210002, China)
Abstract:
The processes
of
culturing CHO(Chinese Hamster Ovary)cells to produce rHuEPO(recombinant
human
Ery
也
ropoietin)were
investigated by using self-prepared
collaεen
c
缸
Tier
,
self-prepared C-DISK carrier
and imported A-DISK carrier
, respectively. The analysis and comparison
of
the anchorage
of
CHO cells,
metabolic rates
of
glucose, expression concentration
of
rHuEPO and the relation between metabolic rate
of
glucose and expression concentration
of
rHuEPO for culturing CHO cells
on
three different carriers were
experimentally carried
ou
t.
The results show that
selιprepared
collagen carrier has the best
bio-activi
句
r
and
highest protein expression in CHO cell culture. The maximum glucose metabolic rate
of
CHO cells cultured on
self-prepared collagen carrier is as
high
出
14
.2
mmol.
mL
-1.h-
1
,and which is 16
.4%
higher than that
on
C-DISK
C
缸
rier
,
12.8% higher than that
on
A-DISK carrier, the final rHuEPO expression concentration in CHO cell
culture cultured
on
selιprepared
collagen carriers reaches 282
U-
mL
-1
after culturing ten days, and it is 30%
higher than that
on
C-DISK carrier, 18% higher than that
on
A-DISK carrier. The
selιprepared
collagen
carrier is a kind
of
high efficient and cheaper carrier which can be used for large-scale animal cell culture. The
experimental results also
showthat
the metabolic rate
of
glucose and expression concentration
of
rH
uE
PO in
CHO cell culture cultured
on
di
岱
rent
carriers are strongly relevan
t,
but they are not in synchronization. This
means
也
at
CHO cell culture stage can be detected and regulated by measuring the change
of
glucose metabolic rate.
Key
words:
cell culture; carrier; CHO cells; recombinant human
Ery
由
opoietin
(rHuEPO)
收稿日期
2004-07-13
,修订日期
2004-11-22.
基金项目
z
国家自然科学基金。
0176018
,
20376034)
,江苏省科技厅资助项目(BK2
003084
和
BS2003030)
,国家"十五"科技攻关项目
(2004B
A3
13BI6).
作者简介=陈英文(1
978-)
,男,山东莱阳人,南京工业大学讲师,硕士.
通讯联系人
z
沈树宝,
E-
ma
i1:
zsbshen
@n
jutedu.cn