Bio-Rad Profinia™ Protein Purification Instrument - Manual (Page 36)

Section 6
Bio-Rad Methods
The Bio-Rad Methods function of the Profinia™ system is used to select preprogrammed
purification and desalting methods. These methods have been optimized on the Profinia
instrument with Profinia purification kits to provide the highest purity and yield. Purification
kits are available for all Bio-Rad methods that have been preprogrammed in the instrument.
When a method type and the associated options are selected, the Profinia screen displays
a method-specific diagram of the system that shows where each cartridge, bottle, and tube
is placed on the instrument. Bottles and cartridges in the purification kits are labeled to
match the screen diagrams.
The purification procedure used in Bio-Rad methods has the following steps:
1. Select method type and options.
2. Select sample flow rate, cartridge wash time, or incubation time.
3. Enter run and sample information.
4. Install reagents, sample, and fraction tubes.
5. Install cartridge(s).
6. Start run.
7. Monitor run.
8. Clean sample and fraction lines.
9. Select and perform end-of-run options.
10. Shut down system or start a new method.
Instrument Operating Temperature
For Bio-Rad methods that include the desalting step, the instrument operating
temperature is critical. Maximum flow rates for the desalting cartridges are lower at 4°C
than at room temperature. Flow rates over maximum limits compress the bed volume and
cause over-pressure errors. Make sure the operating temperature is set to match cold room
or room temperature operating conditions before programming your run. Methods with
desalting steps have longer run times at the cold room setting due to the lower maximum
flow rates. The operating temperature setting displays as a status indicator in the upper
right corner of the screen. If the cold room operating temperature is selected, “C” appears
only when methods are selected that include a desalting step. If the room temperature
setting is selected, “C” will not display. For affinity-only methods, the temperature setting is
irrelevant and will not display. To set the operating temperature:
1. Touch the Data/Utilities button to display the system Utilities screen.
2. Touch the arrow buttons to navigate the list of system utilities; highlight “Select Method
Temperature” then touch Select to display the Select Method Temperature screen
(Figure 6.1).
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