[Comparison of adsorbent with varying arm length and ligand density for the purification of recombinant hepatitis B virus surface antigen].
Novel hydrophobic absorbents were synthesized by immobilizing butyl derivative onto the highly cross-linked agarose beads manufatured in China, which are used as matrix. The effect of the spacer arm length (3C, 8C and 10C) and ligand density (from 13 to 45 micromol/mL) on the hydrophobicity were investigated using purified Hepatitis B surface antigen (HBsAg) expressed by CHO cell lines. Also considering the effects of salt concentration and pH on HBsAg recovery and purification factor, orthogonal experiment design method was used to evaluated the absorbents. The results showed the butyl-S absorbent with the spacer arm length of C8, the ligand density of 22 micromol/mL gel showed the best performance for the separation of HBsAg. Approximately 100% HBsAg recovery and 60 as purification fold were achieved by this media under the operating condition of pH 7.0 and 9% of salt concentrateion.
Li
Rui-Hong
RH
Civil & Environmental Engineering School, University of Science and Technology of Beijing, Beijing 100083, China.
Li
Yan
Y
Bi
Jing-Xiu
JX
Zhao
Lan
L
Zhou
Wei-Bin
WB
Huang
Yong-Dong
YD
Zhang
Yan
Y
Sun
Li-Jing
LJ
Wang
Hua-Jun
HJ
Su
Zhi-Guo
ZG
D000327:Adsorption; D000818:Animals; D016466:CHO Cells; D002847:Chromatography, Agarose / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis / Q000302:isolation & purification*; D057927:Hydrophobic and Hydrophilic Interactions; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification*