Bio-Rad Profinity eXact™ Purification and Tag Cleavage Consumables - Manual (Page 14)

Section 9
Scaling Up
Bio-Scale Mini cartridges are available in 1 ml and
5 ml cartridge formats. The Profinity eXact resin is
also available in 10 ml bottles. For quick scale-up,
two or more cartridges may be connected in series;
backpressure will increase with cartridges in series,
so care should be taken to maintain an overall system
pressure
£45 psi. In addition, Bio-Rad carries an
extensive line of empty chromatography columns
from laboratory scale to process scale. Inquire with
your local Bio-Rad representative or go online to
www.bio-rad.com.
Section 10
Regenerating, Cleaning,
and Storage
Protein cross-contamination, frit clogging, and
increased backpressure may result from cartridge
overuse. It is recommended to dispose of a
23
Fig. 6. Native purification. Purification of Profinity eXact-tagged
Maltose Binding Protein from the soluble fraction using a BioLogic
DuoFlow system. 5 ml of lysate (5 CV) prepared from 50 ml of E. coli
culture was loaded onto a 1 ml cartridge. The cartridge was washed
with 10 CV of bind/wash buffer, and purified protein was eluted with
5 CV of elution buffer following a 30 min cleavage incubation. The
purified product was determined to be 99.5% pure by Quantity
One
® software analysis. Lane 1, Precision Plus Protein unstained
standards; lane 2, soluble lysate; lane 3, flow-through; lane 4, wash;
lane 5, purified product.
0.00
0.25
0.50
0.75
1.00
1.25
1.50
1.75
2.00
AU
10.00 20.00 30.00 40.00
Min.Tenth
Tube #: 1 2 3 4 567 9 1112
22
kD
250
150
100
75
50
37
25
20
15
10
Lane Order
1 2 3 4 5
Cleavage
Incubation
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