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Cell-containing Sampling

Cell-containing Sampling

Cell-containing sampling is very useful for a more detailed analysis of the reactor content. It enables to analyze the cellular material as well as nutrients and metabolites.

This method is particularly suitable for investigating the cell state (cell number, viability, morphology, apoptosis) or after cell disruption for performing further analysis of the cell contents.

The extracted contents of the reactor can be added directly to the downstream analyzers or stored by means of a fraction collector.

 

1. Components

1.1 TRACE C2 Sampling - Automated online Sampling

The TRACE C2 Sampling is a universal online bioreactor sampling system which can automatically withdraw samples from up to four bioreactors. Both cell-containing samples and cell-free samples can be obtained.


1.2. TRACE single-use tubing sets

Depending on the sampling method, sterile tubing sets are available that are easily mounted on the device. They include all components for sampling, such as pumps, valves, filters and the corresponding connector the bioreactor or the sampling probe.

Solutions for rinsing and disinfection are automatically promoted through the tubing system to ensure optimum quality of the individual samples and to exclude contamination of the process.

2. Methods

For the diverse needs of an effective online process monitoring and sample TRACE offers a wide range of methods which can be selected by the user according to the process conditions and measurement tasks.

Currently, the following methods for cell-containing sampling are available:

1.    Cell-containg Sampling
2.    Cell-containg Sampling 4-channel
3.    Cell-containg Sampling Cytometer
4.    Cell-containg Sampling Cytometer 4-channel

2.1 Cell-containing Sampling

Cell-containing sampling is very useful for a more detailed analysis of the reactor content. It enables to analyze the cellular material as well as nutrients and metabolites. This method is particularly suitable for investigating the cell state (cell number, viability, morphology, apoptosis) or after cell disruption for performing further analysis of the cell contents.The extracted contents of the reactor can be added directly to the downstream analyzers or stored by means of a fraction collector.

For installation the bioreactor the sampling line „Sample (in)“ of the sterile tubing set is directly connected to the sampling port of the reactor. For sampling pump 1 automatically draws the required amount of medium in selected intervals or at defined times and releases it via the "Sample (out)" port to the downstream systems. Subsequently, a self-running routine in which the tubing set is cleaned and disinfected with the aid of buffer containing disinfectant. Then the line is flushed with sterile filtered air to prevent carryover of individual samples.

Prior to sampling, the line “Sample (in)” is pre-rinsed with medium to obtain a representative sample. After sampling and between individual samplings the lines are rinsed with buffer ("Buffer") and air ("Air"). Again, this process is automatic. The buffer solution and the medium-rinsing are disposed via the waste line ("Waste").

During longer intervals, the lines remain filled with disinfectant resp. cleaning solution to safely shield the reactor contents against contamination. 3N NaOH, alcohol solutions or TRACE cleaning solution can be used for this purpose.

In addition to the times or intervals of sampling, the volume of the respective sample can be chosen in the range of 0.5 - 20 ml. The individual dead volume resulting from different connectors to the bioreactor, sampling ports or connecting lines can be compensated by selection of a respective purge volume.

Settings:
- Sample volume
- Purge volume
- Interval every x minutes, or the time of sampling

 

2.2 Cell-containing Sampling 4 Channel

In addition to the method described in section 2.1, a 4-channel version of the tubing set can be used, in which up to 4 different reactors can be operated by TRACE C2 Sampling. The particular reactor is then switched on via a selector valve, and the probes are activated one after another or at set points in time.
Settings (for each reactor individually):
- Sample volume
- Purge volume
- Interval every x minutes, or the time of sampling

 

2.3 Cell-containing Sampling Cytometer

In addition to the method described in section 2.1, a version of the tubing set can be used, in which a cytometer is bypassed to the sample (out) line. During the sampling process, a second pump withdraws a small portion of the sample for parallel analysis with the aid of a flow-through Cytometer („Cytometer (out)“).

 

 

 

The flowrate of this bypass can be set in the range of 5 – 30 µl/min.
Settings:
- Sample volume
- Purge volume
- Flow rate of bypass to Cytometer
- Interval every x minutes, or the time of sampling

 

2.4 Cell-containing Sampling Cytometer 4 Channel

In addition to the method described in section 2.3, a 4-channel version of the tubing set can be used, in which up to 4 different reactors can be operated by TRACE C2 Sampling. The particular reactor is then switched on via a selector valve, and the probes are activated one after another or at set points in time.

Settings (for each reactor individually):
- Sample volume
- Purge volume
- Interval every x minutes, or the time of sampling

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