层析原理及工艺开发
主要内容
1. 层析原理 2. 蛋白纯化工艺开拓
1/ GE Title or job number / 10/15/2008
2/ GE Title or job number / 10/15/2008
Principles of operation for chromatography techniques
Chromatography techniques
.Terms
and Parameters
Gel filtration
Ion exchange
Hydrophobic interaction
Affinity
Reversed phase
3/ GE Title or job number / 10/15/2008
Separation volumes
Vo VR Vt Vi Vc = = = = = void volume retention volume or elution volume total liquid volume of the bed inner pore volume = Vc - Vs - Vo total geometric volume of the column
Efficiency
AU280
N = 5.54 (
Wh = peak width at half peak height
Vr 2 ) Wh
void volume Vo
N = Number of theoretical plates test: 1% solution of acetone – about 0.5% of column volume, 0.2 AUFS at 280 nm. alternatively use 2 M NaCl and conductivity monitor
VR
volume of the gel matrix Vs
Vo
Vt
pore volume Vi
Vc
Vr
5/ GE Title or job number / 10/15/2008
6/ GE Title or job number / 10/15/2008
Symmetry: the shape of your peak
AU280
Linear Flow Rate b a
Volume Flow rate (ml/min) x 60
1 is perfect shaped Gauss curve 1 peak is tailing 1 peak is leading
As =
As =
ab

下一页