Bio-Rad Profinia™ Protein Purification Instrument - Manual (Page 75)

Section 7
Program Methods
The program methods function of the Profinia™ system is used to create methods by editing
the Bio-Rad purification and desalting method templates. The method template remains the
same in terms of number and sequence of steps; however, step parameters such as flow
rate, column volume, and peak detection can be edited to create a program method. Once
a method is programmed it can be stored in memory for future access and reuse.
The purification procedure used in program methods has the following steps:
1. Select method type and options.
2. Enter run and sample information.
3. Edit method; save method.
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 procedures.
10. Shut down system.
Instrument Operating Temperature
For program 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 tem-
perature. 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. See section 6.2 for flow rate details, or section 9.3.13 for information on
setting the operating temperature.
Warning: When operating at 4°C, make sure that selected methods that include desalting
are programmed for 4°C. Methods created using room temperature operating parameters
could have flow rates that are too high for 4°C and damage to the cartridges can occur.
Note: Do not change the instrument operating temperature setting to cold room (indicated
by a “C” in the upper-right of the touch screen) when using the cooling accessory. Keep the
operating temperature setting at room temperature unless the entire instrument is operating
in a cold room environment.
Figure 7.1 shows the screen sequence when starting a program method.
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