Peptide-MHCI complex preparation

Eureka Therapeutics has developed a method for the generation of peptide-MHC complexes with high fidelity.

The complexes are folded in the presence of humanβ-2 microglobulin, a chaperone that ensures correct conformation. After purification of complexed monomers using size exclusion chromatography, the complexes are biotinylated and re-purified by anion-exchange chromatography.

Adaptive library panning for human antibodies (ALPHA)

Eureka’s proprietary E-ALPHA® phage display platform is designed specifically for fully-human antibody drug discovery against intracellular cancer antigens.

The platform generates highly-specific, high-affinity antibody candidates against both conventional and novel human targets. The platform’s core component is a fully-human phage library containing 100 billion unique human antibody fragments. The library is one of the largest in the world, and is comprised of multiple sublibraries:

  • Naïve Human Libraries (scFv and Fab Formats)

  • Semi-Synthetic Human Libraries (scFv and Fab Formats)

  • Naïve Human Libraries from Disease Patient PBMCs (scFv and Fab Formats)
Constructed from PBMCs isolated from patients with various diseases, including autoimmune.


High-throughput screening for specificity

Each cell in our body expresses an average of 500,000 different MHCI/peptide complexes.

It is essential for an antibody immunotherapy to be able to distinguish between the “target” MHCI/peptide complex versus the MHCI molecule itself or an MHCI molecules bound to an “off-target” peptide. At Eureka we have developed a novel high-throughput screening platform for assessing specificity, using cell arrays expressing MHCI/peptide complexes and recombinant protein arrays. This platform enables us to exclude drug candidates with undesired cross-reactivity at early stages of drug discovery, and ensures that our CAR/TCR based therapies are exquisitely precise in binding their designated target.

In-depth experience evaluating antibody candidates

Eureka Therapeutics has successfully isolated highly specific candidates across a broad range of disease targets.

A few highlights from our analytical tool box:

  • Epitope mapping by scanning mutagenesis

  • Binding analysis to HLA subtypes

  • Engineering hits into various antibody formats for analysis and testing:

1. Human & mouse IgG subclasses

2. Fc glyco-engineering

3. Anti-CD3 bispecific formats

  • Crystalization of mAb/MHCI-peptide complex to discern structure and binding interface

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