60
The recommended cycle will require custom programming to incorporate a 90
minute preheat and 2-3 moderated prevac pulses to control the rate of temperature
change within the cartridge to a rate of 1°C/min during the warm up. Repligen
recommends that during the cycle development, several thermocouples should be
positioned within the assembly to track the rate of temperature change and confirm
that all positions are at 121-123°C for the duration of the sterilization section of the
cycle.
8.2.2 While autoclave conditions may vary based on requirements, the following are typical
autoclave conditions for the Filtration Assembly. Please refer to the XCell™ ATF System
Filter Preparation and Autoclave Guide for more details. The entire cycle may take 4-5
hours.
• The assembly should be placed in the autoclave at a 45° angle and be allowed to
warm up for a period not less than 60 minutes with steam in the jacket, but not
in the chamber.
• The active cycle will begin with a 5-minute purge where steam flows through the
autoclave directly to the drain with minimal increase in chamber pressure.
• Secondly, there are 2-3 vacuum pulses with “hold” periods in between. These
are critical to evacuate all of the air within the assembly and ensure that there is
a uniform temperature throughout the assembly prior to engaging the ramp up
to reach the sterilization temperature.
• Follow with a 1-minute steam purge to about 6psi, and 108
°C
• Ramp up slowly, 1C/min, to sterilization temperature / pressure, of 121 °C - 123
°C /
16psi
• Sterilize for ~ 55-60 min.
• Ramp down slowly, with wet cycle exhaust protocol or 0.5-1
°
C/min to 100
o
°C
• Allow the system to cool to 50 °C before handling
8.2.3 Open autoclave door, inspect the system, re-tighten all clamps (about ¼ turn).
8.2.4 Carefully remove the Filtration Assembly from the autoclave.
8.2.5 Preferably, remove the Filtration Assembly from autoclave into clean area.
8.2.6 Allow system to cool to room temperature in a clean area.
8.2.7 Pressurize system to about 5psi using sterile air source through one of the retentate
vent filters. This step is optional.
8.2.9 After cooling down to room temp or after pressurization of system, close all valves.
1.2.10 The system may be stored until use.
8.3 Sterilization of Filtrate / Harvest line
The Filtrate / Harvest line with end filter will be sterile following autoclave. One must assure
however that the Harvest line has been prepared with the appropriate tubing for use with
the filtrate / harvest pump. One may also prepare the Filtrate / Harvest line, post
autoclaving, using a tube welder. If a tube welder cannot be used, a disposable sterile
coupling may be used.
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