p. 7
7. When the bed ha s stabilized, clo se the bottom out let and stop the pum p.
8. If using a packi ng reservoir, discon nect the reser voir and fit the ad aptor
to the column.
9. With the adap tor inlet disconne cted, push down t he adaptor
approximately 2 mm into t he bed, allowing t he packing solut ion to flush
the adaptor inlet.
10. Connect th e pump, open the bot tom outlet and cont inue packing.
The bed will be furth er compressed at t his point and a space w ill form
between the bed sur face and the adaptor.
11. Close the bot tom outlet. Di sconnect the col umn inlet and lower t he
adaptor approximate ly 2 mm into the bed. Con nect the pump. Th e
column is now ready to use.

3.3 Packing large scale columns

General packing recommendations

Columns can be packed in di fferent ways dep ending on the type of co lumn
and equipment used. Al ways read and follo w the relevant colum n
instruction man ual carefully. For g eneral process-s cale column packin g
instructions, p lease visit supp ort section a t:
www.gehealthcare.com/protein-purifi cation
4. Evaluation of packing
To check the quality of the pack ing and to monitor th is during the work ing
life of the column, column ef ficiency shou ld be tested direc tly after packi ng,
prior to re-use, and whe n separation per formance is see n to deteriorate.
The best method of exp ressing the eff iciency of a packed co lumn is in terms
of the height equivalent t o a theoretical pla te, HETP, and the asymmetr y
factor, AS. These values are e asily determine d by applying a samp le such as
1% acetone solution to the co lumn. Sodium chlo ride can also be use d as a
test substance. Use a con centration of 2.0 M NaCl i n water with 0.5 M NaCl i n
water as eluent.