Phoneutria nigriventer spider can cause severe envenomation in humans principally due to its venom toxin δ-ctenitoxin-Pn2a. Current low yielding antivenom production is extremely complicated and dangerous. Furthermore, δ-ctenitoxin-Pn2a cystine-knot motif provides exceptional stability hampering immune response activation. Here, epitopes from δ-ctenitoxin-Pn2a were identi ed, and antigenic peptides were designed for their potential use in antivenom production. The Immune Epitope Database Analysis Resource was used to identify the G 34 YFWIAWYKLANCKK 48 epitope and used to design antigenic peptides. The Cys was replaced by α-aminobutyric acid (Abu) to avoid disul de bonds formation. To increase their immunogenicity, branched and N-palmitoylated peptides were synthesized.Ac-GYFWIAWYKLAN-Abu-KKG-NH 2 (A), (Ac-GYFWIAWYKLAN-Abu-KK) 2 -KG-NH 2 (B), Palm-GYFWIAWYKLAN-Abu-KKG-NH 2 (C) and (Palm-GYFWIAWYKLAN-Abu-KK) 2 -KG-NH 2 (D) were synthesized using solid-phase peptide synthesis (SPPS) techniques and analyzed by ESI-MS demonstrating their identity. Also, they were evaluated by RP-HPLC, and all the chromatograms showed only one principal peak except that of the N-palmitoylated branched peptide which showed two principal peaks probably due to the presence of two conformations in slow interconversion. Cytotoxicity was evaluated on the murine macrophage cell line RAW264.7 by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay in the presence of increasing doses of each peptide (0.25-10.0 µM). Peptide A did not exhibit cytotoxicity between 0.25-10.0 µM, while B, C and D showed cytotoxicity over 10.0, 5.0 and 2.5 µM respectively. NF-κB cellular distribution was evaluated by immuno uorescence, after exposing macrophages to 0.5 µM of each peptide. An early activation was observed for all the assayed peptides demonstrating that they are promising candidates for their in vivo evaluation as immunogens in antivenom production.
Phoneutria nigriventer spider can cause severe envenomation in humans principally due to its venom toxin δ-ctenitoxin-Pn2a. Current low yielding antivenom production is extremely complicated and dangerous. Furthermore, δ-ctenitoxin-Pn2a cystine-knot motif provides exceptional stability hampering immune response activation. Here, epitopes from δ-ctenitoxin-Pn2a were identified, and antigenic peptides were designed for their potential use in antivenom production. The Immune Epitope Database Analysis Resource was used to identify the G34YFWIAWYKLANCKK48 epitope and used to design antigenic peptides. The Cys was replaced by α-aminobutyric acid (Abu) to avoid disulfide bonds formation. To increase their immunogenicity, branched and N-palmitoylated peptides were synthesized. Ac-GYFWIAWYKLAN-Abu-KKG-NH2 (A), (Ac-GYFWIAWYKLAN-Abu-KK)2-KG-NH2 (B), Palm-GYFWIAWYKLAN-Abu-KKG-NH2 (C) and (Palm-GYFWIAWYKLAN-Abu-KK)2-KG-NH2 (D) were synthesized using solid-phase peptide synthesis (SPPS) techniques and analyzed by ESI-MS demonstrating their identity. Also, they were evaluated by RP-HPLC, and all the chromatograms showed only one principal peak except that of the N-palmitoylated branched peptide which showed two principal peaks probably due to the presence of two conformations in slow interconversion. Cytotoxicity was evaluated on the murine macrophage cell line RAW264.7 by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay in the presence of increasing doses of each peptide (0.25-10.0 µM). Peptide A did not exhibit cytotoxicity between 0.25-10.0 µM, while B, C and D showed cytotoxicity over 10.0, 5.0 and 2.5 µM respectively. NF-κB cellular distribution was evaluated by immunofluorescence, after exposing macrophages to 0.5 µM of each peptide. An early activation was observed for all the assayed peptides demonstrating that they are promising candidates for their in vivo evaluation as immunogens in antivenom production.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.