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Membrane-based cell affinity chromatography to retrieve viable cells

E Mandrusov1, A Houng, E Klein

  • 1Department of Chemical Engineering, Columbia University, New York, New York 10027, USA.

Biotechnology Progress
|March 1, 1995
PubMed
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This study demonstrates a novel cell affinity chromatography (CAC) system for separating and recovering B-cells from mouse spleens. The method uses antibody-coated cellophane and acid elution, achieving viable cell recovery.

Area of Science:

  • Biotechnology
  • Cell Separation
  • Immunology

Background:

  • Cell separation is crucial for research and clinical applications.
  • Existing methods often involve complex procedures or result in cell damage.
  • There is a need for efficient and gentle cell isolation techniques.

Purpose of the Study:

  • To develop and demonstrate a novel cell affinity chromatography (CAC) system.
  • To achieve separation and live recovery of specific cell types from a mixture.
  • To optimize conditions for cell attachment and elution.

Main Methods:

  • Immobilization of anti-murine IgG onto a cellophane support using a carbonyldiimidazole (CDI) linker.
  • Flowing murine splenocytes over the support, allowing B-cells to attach at a shear rate of 15 s-1.

Related Experiment Videos

  • Washing nonspecifically bound cells at 315 s-1 and eluting B-cells using transmembrane diffusion of hydrochloric acid (pH 1) combined with shear flow.

    • Main Results:

    • An average of 250 cells/mm2 attached to the antibody-immobilized surface at 15 s-1.
    • Successfully displaced attached cells using acid diffusion from the opposite side of the membrane.
    • Achieved an average of at least 60% viable B-cell recovery, assessed by Trypan Blue dye exclusion.

    Conclusions:

    • The developed CAC system effectively separates and recovers B-cells from murine splenocytes.
    • The acid-mediated elution method preserves cell viability.
    • This technique offers a promising approach for selective cell isolation.