Fig. 1.

Experimental overview. A. Eukaryotic ribosome display selection and CCK-BR scFv antibody generation. The scFv’s were generated using cell-free ribosome display from mice spleens immunized against a human CCK-BR peptide fragment selected at an extracellular binding region, followed by eukaryotic ribosome display selection. B. Timeline for in vivo model induction, behavioral and downstream tissue analysis. After acclimatization the FRICT-ION or SNI neuropathic pain model was induced in anesthetized mice. Three weeks later when hypersensitivity was stable, the CCK-BR scFv 77-2 was administered intraperitoneally. Hypersensitivity testing with von Frey fibers continued weekly. The cold hypersensitivity was assessed in week 6, and anxiety-, depression-, and cognitive-like behaviors were assessed in Weeks 8–9. Tissues were collected at 10 weeks post-injury for immunohistochemical, Western blot, and RNAseq analysis or 3–4 weeks post-injury electrophysiological analysis.