Authors
Xinying Dong, Huimin Tao, Xiaowei Gao, Hao Song, Jing Li, Tao Wang, Rui Luo, Yuan Sun, Hua-Ji Qiu, Yuzi Luo
Published in
Virology. Pages 111117. Oct 28, 2026. Epub Oct 28, 2026.
Abstract
African swine fever virus (ASFV), the etiological agent of African swine fever, continues to impose a heavy disease burden on the global swine sector. The lack of broadly protective vaccines makes serological surveillance a cornerstone of disease management strategies. Here, we report the first isolation of alpaca-derived nanobodies (Nbs) specifically targeting the ASFV p30 protein. From a phage-displayed immune library, we selected a high-affinity clone (Nb1) and engineered a lentiviral system for stable, secretory production of an Nb1-horseradish peroxidase (Nb1-HRP) fusion protein in suspension-adapted HEK293 cells, incorporating a flexible peptide linker to preserve the respective biological activities of both the Nb and the enzyme. Biolayer interferometry verified that Nb1-HRP bound to p30 with ultrahigh affinity, yielding an equilibrium dissociation constant (KD) of 65.3 pM. The resulting probe enabled the development of a blocking ELISA (bELISA) for detecting anti-ASFV antibodies in pig sera. The bELISA demonstrated superior sensitivity compared with a commercial ELISA kit (ID.vet, France) and exhibited no cross-reactivity with antibodies against other prevalent swine viruses. Parallel testing of 267 field swine serum samples yielded an overall diagnostic agreement of 98.1% (262/267) between the novel bELISA and the commercial kit. Notably, the assay detected seroconversion at 7 days post-infection, three days earlier than the commercial kit. These findings establish this Nb-based platform as a scalable and sensitive alternative for early ASFV serosurveillance.
PMID:
42977606
Bibliographic data and abstract were imported from PubMed on 04 Nov 2026.
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