文档介绍：实验室层析柱装柱技术介绍
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Column packing
好的柱效是层析成功的关键
不同种类的凝胶、不同的层析柱有不同的装柱方法
柱效测定的方法
层析柱的维护
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Column packing
Rabies vaccine
Sepharose 4 Fast Flow
5% column vol sample
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Column packing
ml mAU*ml (volume) % mAU
No Ret Area Area/Peak area Height
1
2
每米塔板数：N=1600
ml mAU*ml (volume) % mAU
No Ret Area Area/Peak area Height
1
2
每米塔板数：N=3500
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Column packing
N/m= (Ve/Vh)2100/L
As = b/a
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Column packing
检测装柱效果一般用<% (30um介质)或<1% (90um介质) 柱体积的1%***测柱效和峰形
The linear flow rate should be 30 cm/h for 34 µm and 50 µm media and 20 cm/h for 90 µm media
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线性流速
对体积不同的柱子来说，体积流速不能直接用作比较，一般都换算成线性流速，再作比较：
体积流速 (ml/min) x 60
线性流速 (cm/hr) = 揪揪揪揪揪揪
柱子横切面积 (cm2)
Column packing
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Column packing
It is important that the column is properly equilibrated before the
packing is evaluated (~ 2 column volumes). It is preferable to run
three test runs to see whether the test values are stable.
If a poor result improves during this test, the reason can be that the
column was not properly equilibrated.
To check that the bed is stable, run the column at 70% packing
pressure for 20 hours and test it again (preferably three test runs).
Note that pressure spikes may cause poor packing (cracking). If
this happens, an air trap and a pressure relief valve are needed
between the pump and the column. The pressure relief valve
should be placed between the air trap and column.
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Column packing
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装柱注意点
选择一根设计良好的柱子
仔细混合均匀凝胶浆
在生产时使用柱子的温度下装柱
用平缓的动作将凝胶浆一次性加入柱子中
用恒压或恒流装柱
稳定和平衡凝胶柱床
检查柱效，必要时重装
Column packing
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