Branch Lab Translational Cell Therapy (Hannover)

Methods

Development of GMP-compliant protocols for the fully automated generation of CAR-expressing donor NK cells

GMP-compliant cell separation

CliniMACS Plus, Miltenyi Biotec (separation)

GMP-compliant cell separation

CliniMACS Prodigy, Miltenyi Biotec (separation / expansion)

GMP-suitable quality control using 10-color flow cytometry

Navios (Beckman Coulter)

ADCC (“antibody-dependent cell-mediated cytotoxicity”)

GMP-suitable accumulation (“GMP sorting”) of the CD16-positive (cytotoxic) subpopulation of activated donor NK cells used in combination with IgG antibodies to specifically eliminate cancer cells.

GMP-compliant cell separation

CliniMACS Prodigy (Miltenyi Biotec)

GMP-suitable quality control using 10-color flow cytometry

Navios (Beckman Coulter)

GMP-suitable "sorting"

MACS Quant Tyto (Miltenyi Biotec)

8-color fluorescence microscopy (Olympus) with quantitative quality control based on IX81 ScanR analysis software for developing potency assays that can be validated

IX81 (Olympus) ScanR fluorescence microscope
© Fraunhofer IZI
IX81 (Olympus) ScanR fluorescence microscope.
CAR-mediated "retargeting" of specific effector-target interactions
© Fraunhofer IZI
CAR-mediated "retargeting" of specific effector-target interactions.
Time-lapse imaging and tracking of CAR-modified donor NK cells (green/yellow) against resistant AML blasts (blue)
© Fraunhofer IZI
Time-lapse imaging and tracking of CAR-modified donor NK cells (green/yellow) against resistant AML blasts (blue).

GMP-compliant electroporation (transfection) of activated donor NK cells in a closed tubing set system as an alternative to transduction with alphaviral / lentiviral vectors

Maxcyte GT electroporator (Maxcyte)
© Fraunhofer IZI
Maxcyte GT electroporator (Maxcyte).
Electroporation without mRNA: Flow cytometry (Navios) to the left, fluorescence microscopy (Olympus IX81) to the right
© Fraunhofer IZI
Electroporation without mRNA: Flow cytometry (Navios) to the left, fluorescence microscopy (Olympus IX81) to the right.
Electroporation using GFP-tagged mRNA
© Fraunhofer IZI
Electroporation using GFP-tagged mRNA.