The novel Fab-TACS® technology for Cell Separation
Fab-TACS® - Traceless Affinity Cell Selection - technology represents an affinity chromatography system for non-magnetic isolation of immune cells. It is based on Twin-Strep-tagged Fab fragments, which reversibly capture and release the target cells. The Fab-TACS® technology delivers label-free, non-activated cells suitable for immunological or cell biological assays.
Fab-TACS® technology perfectly combines the benefits of positive and negative cell separation and therefore allows the purification of:
- Highly viable cells
- Highly pure cells directly from whole blood
- Label-free cells due to low affinity Fabs
Automated and Manual Cell Separation based on the Fab-TACS® technology
|Manual Cell Separation||Automated Cell Separation|
The Fab-TACS® Gravity column is a chromatography column filled with a Strep-Tactin® coated matrix made of cell-grade agarose. It allows an easy and specific separation of target cells in high yields and purity directly from whole blood, buffy coat, mouse spleen or other single-cell tissue suspensions.
The FABian® cell selection device is a fully automatic bench top instrument which automates the entire selection procedure based on IBA's novel Fab-TACS® technology. The cells of interest can be isolated in high yields, high viability and purity from diverse sources such as whole blood, buffy coat, mouse spleen cells or other single-cell suspensions.
Fab-TACS® technology for Fab-based Traceless Affinity Cell Selection
A standard problem in cell preparations is limited purity and
recovery, which causes variability in quality and potency of cell products.
Highly pure populations can be obtained best using positive separation
techniques. However, the latter are often based on high affinity antibodies
whose usage causes unfavorable effects like strong and almost irreversible
binding to cells, cell stimulation as well as receptor blockade.
IBA’s Strep-tag®/Strep-Tactin® system is the platform for
the novel traceless affinity cell selection (Fab-TACS®) of cells. Fab-TACS® enables cell separation without the need for high affinity
antibodies and ferromagnetic beads. In contrast, Fab-TACS® uses an
immune-affinity chromatography based on Strep-tagged Fab-fragments (Fab-Streps)
for reversible capture and release of target cells.
In more detail, a Strep-Tactin® coated matrix made of cell-grade agarose specifically binds low affinity Fab fragments. However, multiple Fab-Streps on IBA’s unique agarose bind target cells with high avidity. Hence, if a single-cell suspension passes through the agarose matrix target cells adhere to the matrix based on the exclusive binding of the Fab-Strep to the target cell. Non-target cells are efficiently washed away. In a final step, the addition of biotin causes the elution of the target cells and the Fab-Streps spontaneously self-dissociate from the cell surfaces due to their reduced affinity.
Thus, the innovative Fab-TACS® procedure delivers label-free, non-activated target cells in a standardized manner of highly reproducible quality from a variety of single cell suspensions, e.g. whole blood or buffy coat allowing a multitude of downstream applications.
General procedure of Fab-TACS® - Workflow
Columns for either Fab-TACS® Gravity or FABian® are filled with a Strep-Tactin® coated matrix made of cell-grade agarose. Strep-tagged Fab fragments (Fab-Streps) specifically bind to the matrix. Subsequently, whole blood or samples from other sources, e.g. mouse spleen cells pass through the column. Target cells adhere to the matrix based on the exclusive binding of the Fab-Strep to the target cell. Non-target cells are efficiently washed away. In a final step, the addition of biotin causes the elution of the target cells and the Fab-Streps. After elution, the Fab-Streps self-dissociate from the cell surfaces. The label-free authentic target cells are now ready for further downstream applications.
Further details can be found here: