GE 71-5000-15 AD - Manual (Page 7)

p. 7
7. When the bed has stabilized, close the bottom outlet and stop the pump.
8. If using a packing reservoir, disconnect the reservoir and fit the adaptor
to the column.
9. With the adaptor inlet disconnected, push down the adaptor
approximately 2 mm into the bed, allowing the packing solution to flush
the adaptor inlet.
10. Connect the pump, open the bottom outlet and continue packing.
The bed will be further compressed at this point and a space will form
between the bed surface and the adaptor.
11. Close the bottom outlet. Disconnect the column inlet and lower the
adaptor approximately 2 mm into the bed. Connect the pump. The
column is now ready to use.
3.3 Packing large scale columns
General packing recommendations
Columns can be packed in different ways depending on the type of column
and equipment used. Always read and follow the relevant column
instruction manual carefully. For general process-scale column packing
instructions, please visit support section at:
www.gehealthcare.com/protein-purification
4. Evaluation of packing
To check the quality of the packing and to monitor this during the working
life of the column, column efficiency should be tested directly after packing,
prior to re-use, and when separation performance is seen to deteriorate.
The best method of expressing the efficiency of a packed column is in terms
of the height equivalent to a theoretical plate, HETP, and the asymmetry
factor, A
S
. These values are easily determined by applying a sample such as
1% acetone solution to the column. Sodium chloride can also be used as a
test substance. Use a concentration of 2.0 M NaCl in water with 0.5 M NaCl in
water as eluent.