Use of Taguchi's methods as a basis to optimize hybridoma cell line growth and antibody production in a spinner flask

被引:13
作者
Kallel, H
Zaïri, H
Rourou, S
Essafi, M
Barbouche, R
Koussay, D
Fathallah, DM
机构
[1] Inst Pasteur, Immunol Lab, Mol Biotechnol Grp, Tunis 1002, Tunisia
[2] Inst Pasteur, Viral Vaccines Res & Dev Unit, Tunis 1002, Tunisia
关键词
experimental design; hybridoma; monoclonal antibody; production; Taguchi;
D O I
10.1023/A:1022437514334
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Taguchi's methods were used for the design of an experimental strategy aimed at optimizing cell density and monoclonal antibody (mAb) production from a spinner flask hybridoma culture. 23G11 is an antibody to the human leukocyte adhesion molecule, CR3 or beta2 integrin (CD11b/CD18). It recognizes specifically the A-domain of the alpha subunit CD11b. Anti beta 2 integrin monoclonal antibodies hold a great potential for preventing inflammation mediated tissue injuries. An L8 orthogonal experimental design was used to investigate four different culture components: stirring speed, nature of serum, concentration of serum and nature of media (RPMI 1640 or RPMI 1640 supplemented with glucose and glutamine). The experiments were conducted using two levels for each factor studied and a direct ELISA test was used to estimate the level of antibody production. Statistical analysis of the collected data pointed to the stirring speed and serum concentration, and the interaction between these parameters, as the components that affected cell growth. Antibody production was affected by these factors and by the nature of medium but also by the following two interactions: stirring speed/nature of serum and stirring speed/concentration of serum. This study emphasizes the value of using Taguchi's methods as a basis for optimization of mAb production from a hybridoma culture, in cost effective and significantly less labor intensive ways.
引用
收藏
页码:9 / 14
页数:6
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