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Updated: Oct 29, 2025
Bacterial Inner-membrane Display for Screening a Library of Antibody Fragments
Published on: October 15, 2016
Selection and structural characterization of anti-TREM2 scFvs that reduce levels of shed ectodomain
Aleksandra Szykowska1, Yu Chen2, Thomas B Smith3
1Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK.
Abstract:
Mutations in TREM2, a receptor expressed by microglia in the brain, are associated with an increased risk of neurodegeneration, including Alzheimer's disease. Numerous studies support a role for TREM2 in sensing damaging stimuli and triggering signaling cascades necessary for neuroprotection. Despite its significant role, ligands and regulators of TREM2 activation, and the mechanisms governing TREM2-dependent responses and its cleavage from the membrane, remain poorly characterized. Here, we present phage display generated antibody single-chain variable fragments (scFvs) to human TREM2 immunoglobulin-like domain. Co-crystal structures revealed the binding of two scFvs to an epitope on the TREM2 domain distal to the putative ligand-binding site. Enhanced functional activity was observed for oligomeric scFv species, which inhibited the production of soluble TREM2 in a HEK293 cell model. We hope that detailed characterization of their epitopes and properties will facilitate the use of these renewable binders as structural and functional biology tools for TREM2 research.
Insights
Researchers developed novel antibody fragments targeting TREM2 (triggering receptor expressed myeloid cells 2) to aid neurodegenerative disease research. These tools help study TREM2
Area of Science:
- Neuroscience and Immunology
- Molecular Biology
Background:
- Mutations in TREM2 (triggering receptor expressed myeloid cells 2) are linked to neurodegenerative diseases like Alzheimer's.
- TREM2 is crucial for microglial neuroprotection, sensing damage and initiating signaling, but its activators and regulation are unclear.
Purpose of the Study:
- To generate novel antibody fragments targeting human TREM2.
- To characterize these fragments as tools for structural and functional studies of TREM2.
Main Methods:
- Phage display was used to generate single-chain variable fragments (scFvs) against the human TREM2 immunoglobulin-like domain.
- Co-crystal structures determined scFv binding epitopes.
- Functional assays assessed the activity of scFv oligomers, including inhibition of soluble TREM2 production in a HEK293 cell model.
Main Results:
- Two scFvs were identified, binding to a TREM2 epitope distinct from the ligand-binding site.
- Oligomeric forms of the scFvs showed enhanced functional activity.
- These oligomers inhibited the shedding of soluble TREM2.
Conclusions:
- Novel scFvs targeting TREM2 have been developed.
- These scFvs, particularly in oligomeric form, can modulate TREM2 function by inhibiting its cleavage.
- The characterized scFvs serve as valuable tools for advancing TREM2 research in neurodegeneration.
