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Human T Cell Selection, Activation, and Expansion Kit

Improve Expansion for Healthier Cells

Using BACS™ microbubble technology, the Human T Cell Selection, Activation and Expansion Kit was designed to gently isolate highly pure T cells and activate them for use in cell therapy workflows. In this positive selection workflow, T cells are isolated, activated, and expanded using Akadeum’s Selection, Activation & Expansion Microbubbles in combination with an antibody cocktail that targets CD3 and CD28. This highly scalable process yields large numbers of activated and transducible cells needed to support research and process development in T cell based therapies. The gentleness of flotation-based separation and activation results in fewer exhausted cells compared to legacy activation methods – better research and process development outcomes.

  • Greater activation and lower cellular exhaustion for healthier cells, enabling better therapies
  • Improved expansion of highly activated effector T cell populations.
  • Scalable: process billions of cells with the Alerion™ Microbubble Cell Separation System
  • Closed-system in-bag workflows available
  • GMP and Clinical Ready Grades available to support each stage of your cell therapy pipeline

Product Details

How it Works:

Akadeum’s novel approach to positive selection for CAR-T cell production provides improvements over current industry standard methods, and the innate scalability of BACS™ microbubbles allows for rapid optimization of cell therapy workflows.

Using washed leukopak material or PBMCs as a starting material, T cells are labeled with biotinylated antibodies targeting CD3 and CD28. BACS™ Selection, Activation & Expansion Microbubbles are mixed into the sample where they bind to the labeled cells and gently float them to the surface. Unwanted cells can be removed from below the microbubble layer, and the positively selected T cells transferred to a culture vessel.

BACS™  are inherently inert and do not impact culture conditions. Over time and with cell division, cells gently settle to the bottom of the plate where they continue to expand until collection.

selection activation expansion workflow

 

Additional Information:

  • Compatible with the Alerion™ Microbubble Cell Separation System
  • Antibody Cocktail (biotinylated): CD3 and CD28
  • Storage: 4°C
  • Cell Type: T Cells
  • Sample Species and Source: Human PBMC or platelet washed Leukopak material
  • Cell Separation Method: Positive Selection
  • Capacity: 800 x 106 nucleated cells

This Kit Comes With:

  • Selection Activation & Expansion Microbubbles
  • Selection Activation & Expansion Antibody Cocktail

View the Checklist for Using Akadeum Cell Therapy Kit Products

Key Features

Go from research scale to commercialization with microbubbles. Just use more or less to suit your research needs and keep performance maintained.

scalability

Experimental Overview:

T cells were positively selected from PBMC input material at 100 million total cells (5 mL tube), 1 billion total cells (a 150 mL bag), or 5 billion total cells (BioRise™ Consumable and Alerion™ Microbubble Cell Separation System) following the Human T Cell Selection, Activation, and Expansion protocol.

A) Estimated time based on protocol steps (mixing, incubation, separation) for separation completed in each vessel. As cell numbers increase the workflow time does not lengthen.

B) Comparison of cell viability results at day 1 post selection for cells separated in 5 mL tube, 150 mL bag, and BioRise™ Consumable. All protocols resulted in high cell viability.

C) Total cell expansion results comparing the BioRise™ Consumable selected cells to 5mL tube and 150mL bag vessel controls. Five million captured T Cells were seeded for expansion using G-Rex® 6M Well Plates and complete ExCellerate™ media supplemented with 2% human serum. Additionally, 5U/mL IL2, 5ng/mL IL7, and 5ng/mL IL15 was added on days 0, 4, and 7 of culture. No significant differences were observed in growth rates between the different isolation vessels.

 

The Right Products to Support Your Pipeline

The Human T Cell Selection, Activation and Expansion Kit is ideal for research through process development, with GMP and Clinical Ready versions available to support preclinical to commercialization of cell therapy workflows.

 

Data

More Activation, Less Exhaustion. Healthier Cells.

Optimize results while minimizing potential cellular stress or damage.

 

 

2 x 108 PBMCs were used as starting material for the Akadeum  Human T Cell Selection, Activation, and Expansion kit workflow. Post selection, 5 x 106 T cells/well were seeded and cultured in G-Rex® 6M well plates (Wilson Wolf Manufacturing). Magnetic particles (following the manufacturer’s recommended protocol) were used for comparative T cell selection and were seeded similarly at 5 x 106  per well. Cells for control wells were isolated via Akadeum’s Human T Cell Leukopak Isolation Kit (negative selection) and seeded at 0.5 x 106 per mL in standard 24-well tissue culture plates. Complete ExCellerate™ Media (Bio-Techne), supplemented with 2% human serum (Access Biologicals) was used for all conditions. Additional IL-2, IL-7, and IL-15 cytokines (PeproTech), were added for Akadeum Human T Cell Selection, Activation, and Expansion kit wells while magnetics and control wells were supplemented with IL-2. Harvest timepoints for all conditions were on days 4, 7, and 11.

Resources

User Guide

View the Human T Cell Selection, Activation, and Expansion User Guide
For processing on the Alerion™ Microbubble Cell Separation System, please refer to:
AlerionTM Human T Cell Selection, Activation, and Expansion Kit User Guide

App Notes

T Cell Isolation and Selection from a Diseased Patient Sample: A Comparison Between BACS and Magnetics

Unmatched Scalability, from Process Development to Manufacturing, Akadeum has your BACS

Publications & References

Akadeum Microbubbles: With You from Research to Commercialization

Human T Cell Selection, Activation, and Expansion Kit
AAI 2023 Poster: Novel Workflow for Activation and Expansion of T Cells Using Microbubble Technology
ASGCT 2023 Poster: Buoyant Microbubble-Based T Cell Activation Results in Significantly Reduced Exhaustion Profile

Video

Follow along with our expert-in-residence, Jason Ellis, PhD, as he demonstrates the protocol for using Akadeum’s Human T Cell Selection, Activation, and Expansion Kit with PBMCs and apheresis materials.

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