Single-Dose P2 X4R Single-Chain Fragment Variable Antibody Permanently Reverses Chronic Pain in Male Mice

International Journal of Molecular Sciences

Sabrina L. McIlwrath¹  Marena A. Montera²  Robyn Bartel²  Karin N. Westlund²  Aleyah E. Goins²  Sascha R. A. Alles²  Adinarayana Kunamneni³  Ravi V. Durvasula³  Nikita Suri⁴ 

[1] Biomedical Laboratory Research & Development (121F), New Mexico VA Health Care System, Albuquerque, NM 87108, USA;Department of Anesthesiology & Critical Care Medicine, University of New Mexico Health Sciences Center, Albuquerque, NM 87131, USA;Department of Medicine, Mayo Clinic, Jacksonville, FL 32224, USA;Ross University Medical Center, Miramar, FL 60153, USA;

关键词: small antibody;    chronic pain;    nerve injury;    neuropathy;    hypersensitivity;    anxiety;   

DOI  :  10.3390/ijms222413612

来源: DOAJ

【 摘 要 】

Non-opioid single-chain variable fragment (scFv) small antibodies were generated as pain-reducing block of P2X4R receptor (P2X4R). A panel of scFvs targeting an extracellular peptide sequence of P2X4R was generated followed by cell-free ribosome display for recombinant antibody selection. After three rounds of bio-panning, a panel of recombinant antibodies was isolated and characterized by ELISA, cross-reactivity analysis, and immunoblotting/immunostaining. Generated scFv antibodies feature binding activity similar to monoclonal antibodies but with stronger affinity and increased tissue penetrability due to their ~30% smaller size. Two anti-P2X4R scFv clones (95, 12) with high specificity and affinity binding were selected for in vivo testing in male and female mice with trigeminal nerve chronic neuropathic pain (FRICT-ION model) persisting for several months in untreated BALBc mice. A single dose of P2X4R scFv (4 mg/kg, i.p.) successfully, completely, and permanently reversed chronic neuropathic pain-like measures in male mice only, providing retention of baseline behaviors indefinitely. Untreated mice retained hypersensitivity, and developed anxiety- and depression-like behaviors within 5 weeks. In vitro P2X4R scFv 95 treatment significantly increased the rheobase of larger-diameter (>25 µm) trigeminal ganglia (TG) neurons from FRICT-ION mice compared to controls. The data support use of engineered scFv antibodies as non-opioid biotherapeutic interventions for chronic pain.

【 授权许可】

Unknown