Selection and structural characterisation of anti-TREM2 scFvs that reduce levels of shed ectodomain

Szykowska, A.; Chen, Y.; Smith, T. B.; Preger, C.; Yang, J.; Qian, D.; Mukhopadhyay, S.; Wigren, E.; Neame, S. J.; Gräslund, S.; Persson, H.; Atkinson, P. J.; Di Daniel, E.; Mead, E.; Wang, J.; Davis, J. B.; Burgess-Brown, N. A.; Bullock, A. N.

doi:10.1101/2021.02.07.430123

bioRxiv · 10.1101/2021.02.07.430123

Selection and structural characterisation of anti-TREM2 scFvs that reduce levels of shed ectodomain

Szykowska, A.·Chen, Y.·Smith, T. B.·Preger, C.·Yang, J.·Qian, D.·Mukhopadhyay, S.·Wigren, E.·Neame, S. J.·Gräslund, S.·Persson, H.·Atkinson, P. J.·Di Daniel, E.·Mead, E.·Wang, J.·Davis, J. B.·Burgess-Brown, N. A.·Bullock, A. N.

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Abstract

Single point mutations in TREM2, a receptor expressed by microglia in the brain, are associated with an increased risk of neurodegeneration including Alzheimers disease. Numerous studies support a role for TREM2 in sensing damaging stimuli and triggering signalling 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 characterised. Here, we present phage display generated scFv antibody binders to human TREM2 ectodomain. Cocrystal structures revealed the binding of two scFvs to an epitope on the globular 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 characterisation of their epitopes and properties will facilitate the use of these renewable binders as structural and functional biology tools for TREM2 research.

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Szykowska, A., Chen, Y., Smith, T. B., Preger, C., Yang, J., Qian, D., Mukhopadhyay, S., Wigren, E., Neame, S. J., Gräslund, S., Persson, H., Atkinson, P. J., Di Daniel, E., Mead, E., Wang, J., Davis, J. B., Burgess-Brown, N. A., Bullock, A. N.. 2021-02-08. Selection and structural characterisation of anti-TREM2 scFvs that reduce levels of shed ectodomain. https://doi.org/10.1101/2021.02.07.430123

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