Optimization of expression conditions for soluble protein by using a robotic system of multi-culture vessels.
We have developed a robotic system for an automated parallel cell cultivation process that enables screening of induction parameters for the soluble expression of recombinant protein. The system is designed for parallelized and simultaneous cultivation of up to 24 different types of cells or a single type of cell at 24 different conditions. Twenty-four culture vessels of about 200 ml are arranged in four columns x six rows. The system is equipped with four independent thermostated waterbaths, each of which accommodates six culture vessels. A two-channel liquid handler is attached in order to distribute medium from the reservoir to the culture vessels, to transfer seed or other reagents, and to take an aliquot from the growing cells. Cells in each vessel are agitated and aerated by sparging filtered air. We tested the system by growing Escherichia coli BL21(DE3) cells harboring a plasmid for a model protein, and used it in optimizing protein expression conditions by varying the induction temperature and the inducer concentration. The results revealed the usefulness of our custom-made cell cultivation robot in screening optimal conditions for the expression of soluble proteins.
Ahn
Woo-Sung
WS
Life Sciences Division, Korea Institute of Science and Technology, Seoul 136-791, Korea.
Ahn
Ji-Young
JY
Jung
Chan-Hun
CH
Hwang
Kwang Yeon
KY
Kim
Eunice Eunkyeong
EE
Kim
Joon
J
Im
Hana
H
Kim
Jin-Oh
JO
Yu
Myeong-Hee
MH
Lee
Cheolju
C
D049109:Cell Proliferation; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D012371:Robotics; D013696:Temperature
D011994:Recombinant Proteins