GeneMedi Immuno-X TCE & NKCE Solution

1.1 Effector Cell Isolation — Keep T Cells Resting

SOLIDEX^®-ISOEx Human T Cell Isolation Kits & Nanobeads cover Pan-T, CD4⁺ and CD8⁺ cells, compatible with fresh/frozen PBMCs, whole blood/buffy coat and leukapheresis. Isolation is completed in 30–45 min with >95% purity and a low CD69/CD25 baseline — avoiding the pre-activation caused when positive-selection antibodies crosslink the TCR complex, so a candidate's activation signal is not buried in background.

1.2 Subset & Activation Profiling

SOLIDEX^®-CDEx multicolor antibodies: CD3E/CD4/CD8 for subset gating; CD69 (4–6 h), CD25, CD137 (4-1BB) and OX40 (24–48 h) for time-phased activation; PD-1/TIM-3/LAG-3/TIGIT for exhaustion; target-specific antibodies with calibrated-bead ABC conversion to lock target antigen density before functional assays.

1.3 Function & Potency Assays

• Target-dependent cytotoxicity: luciferase-labeled target cells as primary screen, confirmed by CellTrace Violet + counting-bead absolute counting; outputs EC₅₀, Emax and T₅₀.
• Degranulation and effector molecules: CD107a paired with intracellular perforin/granzyme B to close the “degranulation → killing” chain.
• Cytokines and CRS risk: IL-6, TNF-α, IFN-γ, IL-2 and GM-CSF ELISA, aligned with the 4–13 factor CRS cascade.
• Functional durability: multi-round rechallenge with expansion tracking each round, supported by recombinant human IL-2, IL-7, IL-15 and IL-21.

1.4 Benchmark, PK & ADA

Research-grade Blinatumomab (CD19×CD3) serves as positive control and historical-database benchmark (mean ± 2SD). PK uses a double-sandwich strategy with anti-G4S linker capture plus anti-CD3 / anti-Fc / anti-idiotype detection; ADA follows the four-tier screen–confirm–titer–neutralization workflow.

TCE Solution Detail

2.1 Effector Cell Isolation — Preserve the CD16A Epitope

SOLIDEX^®-ISOEx Human NK Cell Isolation Kits & Nanobeads yield CD3⁻CD56⁺ NK cells, including the CD56^dim CD16⁺ cytotoxic subset. For NKCE, negative selection is a requirement rather than a preference: anti-CD16 positive-selection beads occupy the very binding site most NKCEs target, and crosslinking triggers pre-activation and CD16 shedding. Use ≥3 donors and record CD16A 158V/F genotype to control donor-to-donor functional variability.

2.2 Subset & Phenotype Analysis

SOLIDEX^®-CDEx antibodies cover CD56 (NCAM1), CD16, NKp46 and NKG2D (subsets and activating receptors), NKG2A/KIR (inhibitory background), plus CD69 and CD107a (LAMP-1). CD16 surface expression is monitored at multiple time points to distinguish “molecule inactive” from “receptor shed”.

2.3 Function & Potency Assays

• Primary readouts are CD107a degranulation (4–6 h) and CD69 (4–24 h): no validated Jurkat-based reporter assay applies to NK cells, so functional readouts rely on primary NK flow cytometry.
• Target-dependent cytotoxicity: luciferase primary screen confirmed by absolute-count flow cytometry, with antigen-negative target cells and a low E:T (1:1) condition run in parallel.
• Fratricide control (mandatory): NKCE + NK without target cells to quantify NK-to-NK killing and degranulation background, verifying that the free molecule remains silent.
• Cytokines: NK-derived IFN-γ, TNF-α and GM-CSF are measured and interpreted separately from IL-6 amplification in PBMC-based systems.

2.4 Benchmark, PK & ADA

AFM13 (CD30×CD16A tetravalent TandAb) serves as the methodological benchmark. PK uses anti-G4S linker capture with anti-CD16a / anti-idiotype detection to discriminate intact active molecules from free arms; ADA monitoring is equally essential for Fc-free, non-natural engager formats.