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Chapter 4 Protocol Building Blocks
4
Elutriation
Typical leukaphoresis composition
The distribution of cells based on diameter in a leukapheresis shows quite discrete cell populations or
"fractions". Several important factors need to be considered when developing a protocol to separate
these fractions using elutriation:
• Removal of platelets and debris is readily achieved as part of the PBMC loading step since they are
significantly smaller and will be washed out of the fluidized bed
• The number of cells in each fraction can vary by a significant amount, e.g. some leukapheresis
products have very low monocyte numbers that can make it difficult to establish a monocyte bed.
Red blood cells (RBCs) also vary depending on the donor patient, the process used and the skill of
the leukapheresis operator
• The average size of a lymphocyte is smaller than a monocyte. However, both fractions have a
size distribution, with large lymphocytes overlapping with small monocytes. This overlap in cell
diameter results in a compromise needing to be made between purity and recovery. If you want
high recovery of lymphocytes you will also recover small monocytes. Conversely, to achieve high
purity lymphocytes, you will need to use processing parameters that will not collect the smaller
monocytes which also means you won't collect the larger lymphocytes
80
CTS
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Rotea
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Counterflow Centrifugation System Process Design User Guide
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