Chemokine-binding all-D-CLIPS™ peptides identified using mirror-image phage display

Denisov, S. S.; Bialek, E. L.; Beretta, F.; Smagurauskaite, G.; Ippel, J. H.; Fijlstra, E.; Kale, S. S.; Timmerman, P.; Hackeng, T. M.; Proost, P.; Goldflam, M.; Dijkgraaf, I.

doi:10.1101/2025.09.09.675118

bioRxiv · 10.1101/2025.09.09.675118

Chemokine-binding all-D-CLIPS™ peptides identified using mirror-image phage display

Denisov, S. S.·Bialek, E. L.·Beretta, F.·Smagurauskaite, G.·Ippel, J. H.·Fijlstra, E.·Kale, S. S.·Timmerman, P.·Hackeng, T. M.·Proost, P.·Goldflam, M.·Dijkgraaf, I.

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Abstract

Chemokines are secreted blood proteins, which steer leukocyte migration in the inflammatory response. Neutralization of chemokines is believed to be a beneficial therapeutic strategy for the treatment of inflammation-associated diseases. Proteolytically stable chemokine-binding peptides could be suitable candidates for the development of chemokine-neutralizing agents. Here, we report mirror-image phage display selection of cyclic all-D-peptides against the C-X-C motif chemokine ligand 8 (CXCL8). Selection yielded structurally diverse all-D-peptides with sub-micromolar affinity to the target CXCL8 chemokine and different selectivity to related chemokines. Binding of these all-D-peptides caused dissociation of the native CXCL8 dimer and disruption of its binding to GAGs, without effect on in vitro cell migration. This work demonstrates the example of mirror-image phage display selection of cyclized all-D-peptides and its utility for the development of chemokine-binding agents.

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Denisov, S. S., Bialek, E. L., Beretta, F., Smagurauskaite, G., Ippel, J. H., Fijlstra, E., Kale, S. S., Timmerman, P., Hackeng, T. M., Proost, P., Goldflam, M., Dijkgraaf, I.. 2025-09-10. Chemokine-binding all-D-CLIPS™ peptides identified using mirror-image phage display. https://doi.org/10.1101/2025.09.09.675118

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