IN 45 100 010 • AA • 5 / 12
Scale-up
BabyBio S 1 ml, BabyBio Q 1 ml and BabyBio DEAE 1 ml can be used for purification of up to 50 - 100 mg of
protein. Scale-up from a BabyBio 1 ml column can be carried out by using a BabyBio 5 ml column and applying
a sample volume five times larger. BabyBio columns can be connected in series with a maximum of five columns
(column stacking). This will increase the total capacity of the run accordingly. By connecting BabyBio columns
in series, any column volumes from 1 ml to 25 ml can be obtained. This means a binding capacity of 1000 - 2000
mg of protein can be achieved. Column size selection should be based on estimated amount of target protein
in each run.
Another easy-to-use alternative for scaling up is by using the prepacked OptioBio™ 40S 10x100 or OptioBio 40Q
10x100 columns. OptioBio are prepacked glass columns with WorkBeads 40S or WorkBeads 40Q and with a
column volume of 7.9 ml, see
Related products
.
BabyBio columns are easily connected together without accessories. Up to five columns may be connected in
series (column stacking). The pressure drop across each column bed will be the same as for a single column,
but the upstream columns will be subjected to a higher internal pressure from the added pressure drops from
downstream columns. It may therefore be necessary to decrease the flow rate accordingly in order to avoid
exceed the maximum pressure limit onto the first column. If possible, the maximum pressure of the
chromatography system should be set according to Table 2. Remember always to take the system fluidics
contribution to the pressure into account.
Table 2. Recommended maximum pressure settings for BabyBio columns connected in series. Notice that the maximum pressure over each column is
always 3 bar.
No. of columns in series
Max pressure BabyBio 1 ml
(bar)
Max pressure BabyBio 5 ml
(bar)
1
3.0
3.0
2
6.0
6.0
3
9.0
9.0
4
12
10
1
5
15
10
1
1 The maximum pressure is defined by the column hardware maximum pressure.
Column size selection should be based on the estimated amount of protein to be purified. A test run with a
defined small volume of sample on a BabyBio 1 ml column should be used to estimate the concentration of the
target protein in the sample. A general recommendation is to use 70 - 80% of the column binding capacity. For
large sample volumes with low concentrations of the target protein, it may be suitable to use a larger column
than the one calculated to allow higher sample flow rates, and consequently shorter application time. For
example, using a 5 ml column instead of a 1 ml column allows a flow rate five times higher due to the larger
cross-section of the column. Have in mind that too high flow rate may reduce binding capacity.
For columns larger than 20 ml, it is recommended to pack a single column using bulk resin as the limitations of
column stacking will then impact chromatographic performance. To find out more about Bio-Works bulk
chromatography resins, please visit www.bio-works.com
