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Human T Cell Leukopak Isolation Kit

Negative Selection at Scale

The Human T Cell Leukopak Isolation Kit was developed with BACS™ Microbubbles to isolate untouched T cells from washed leukopak material via negative selection. Leveraging the gentleness of flotation, non-T cells are targeted and removed with antibodies. Negatively selected T cells are suitable for a wide variety of downstream assays, including cell culture, flow cytometry, scRNA-Seq, engineering, or other functional studies. The Human T Cell Leukopak Isolation Kit has the capacity to isolate T cells from an average half-leukopak (12.5 x 10⁹ total cells).

  • Leukopak to purified cells in less than one hour, no matter the cell input number
  • Higher recovery of healthier T cells to enhance CAR-T research
  • Reduce cell impurities and improve your transduction efficiencies
  • Scale up to tens of billions of cells: compatible with the Alerion™ Microbubble Cell Separation System
  • Negative selection workflow eliminates concerns of particles directly bound to T cells

Product Details

How it Works: The Human T Cell Leukopak Biotin Antibody Cocktail is added to the leukapheresis product to label unwanted cell populations. BACS™ Streptavidin Microbubbles are added to the sample and bind to the biotin-labeled cells. Due to their inherent buoyancy, the microbubbles quickly and gently float the non-T cells to the surface. Following centrifugation, the untouched T cells in their original state are collected.

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Additional Information:

  • Compatible with the Alerion™ Microbubble Cell Separation System
  • Antibody Cocktail (biotinylated): CD14, CD16, CD19, CD20, CD36, CD56, CD123, CD235ab.
  • Storage: 4°C
  • Target Cell Type: T Cells
  • Sample Species and Source: Washed Human Leukopak
  • Cell Separation Method: Negative Selection
  • Capacity: 12.5 x 109 total cells

This Kit Comes With:

  • BACS™ Streptavidin Microbubbles
  • Human T Cell Leukopak Biotin Antibody Cocktail
  • Separation Buffer (Ca2+ and Mg2+ free PBS containing 2 mM EDTA, 0.5% biotin-free BSA)
  • Microbubble Separation Tubes*

*If an in-bag process is preferred, please refer to our Human T Cell Leukopak Isolation Kit – GMP User Guide, or contact us.

View the Checklist for Using Akadeum Cell Therapy Kit Products

Key Features

Scalability

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

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Example of scale: 50 million cells in a tube, to 1 billion cells in bag, to 18 billion cells in a BioRise™ Consumable.

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Apheresis material was platelet washed using Thermo Fisher Scientific’s Rotea™ cell washing instrument and then isolated using Akadeum’s negative selection Human T Cell Leukopak Isolation Kit. Samples were processed either in a 50 mL tube at 500 million total starting cells or in a BioRise™ Consumable on the Alerion™ Microbubble Cell Separation System at 5-15 billion cells per consumable. These assays were performed in triplicate.

The Right Products to Support Your Pipeline

The Human T Cell Leukopak Isolation Kit is ideal for research through process development, but rest assured additional grades are and will be available for the remaining stages of the path to commercialization of cell therapy workflows.

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Data

High Purity and Yield at Any Scale

Untouched T cells were isolated from platelet-washed leukopak material using the Human T Cell Leukopak Isolation Kit in accordance with the published protocol.  Isolation was completed with either 100 million cells in a 5 mL tube or 1 billion cells in a 50 mL tube. Samples were run in triplicate. To determine T cell purity and yield, isolated T cells and an aliquot of the initial sample were labeled with CD45-FITC and CD3-APC Fire/750. The fluorescently labeled cells were analyzed by flow cytometry. Dead cells and debris were excluded from the analysis.

t cell isolation process stats
yield t cell isolation stats

Yield and purity for a three-donor leukapheresis sample enriched for T cells with a baseline purity of 41.7% with either the negative selection Human T Cell Leukopak Isolation Kit or a magnetics-based positive selection workflow demonstrates superior performance of Akadeum’s Human T Cell Leukopak Isolation Kit. Data courtesy of third party CDMO.

t-cell isolation

 

Resources

Protocol Sheet

View the Human T Cell Leukopak Isolation Protocol

For in-bag processing, please refer to the:
Human T Cell Leukopak Isolation Kit - GMP Grade User Guide

For processing on the Alerion™, please refer to the:
View the AlerionTM Human T Cell Leukopak Isolation Kit - GMP User Guide

App Notes

High Purity T Cells and Reduced Key Impurities with Akadeum Microbubbles for Improved Efficiencies of Cell Therapy Workflows

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

High Throughput, High Recovery with Akadeum's Microbubble Leukopak Human T Cell Isolation Kit

Publications & References

Starting with RUO? See the GMP kit flyer for your next phase of kit needs.
Product Flyer: Human T Cell Leukopak Isolation Kit - GMP Grade

Akadeum Microbubbles: With You from Research to Commercialization

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