A neutralizing nanobody-based liquid-phase blocking ELISA to assess the protective potency of Senecavirus A vaccine

IF 3.9 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Applied Microbiology and Biotechnology Pub Date : 2025-04-23 DOI:10.1007/s00253-025-13492-4

Suyu Mu, Songjia Pan, Hu Dong, Jinen Wu, Yun Zhang, Shuanghui Yin, Jianing Wang, Tian Wei, Xiaobo Wen, Huichen Guo, Shiqi Sun

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引用次数: 0

Abstract

Senecavirus A (SVA) causes swine blister diseases in the crown of hooves, abortion syndrome, and increased mortality rates in piglets. Vaccination is the best defense against SVA infection. However, evaluations of the efficacy of SVA vaccines currently rely on challenging pigs with SVA, which is time-consuming, labor-intensive, and inconsistent with animal welfare mandates. To establish a replacement for the virus neutralization test (VNT) and SVA challenge tests, we developed a liquid-phase blocking enzyme-linked immunosorbent assay (nbLPB-ELISA) based on V1-VHH as the coating antibody (Ab) and biotinylated V1-VHH as the detection Ab. Under optimized conditions, the VNT and nbLPB-ELISA results were strongly correlated (Pearson R² = 0.84; p < 0.00001). Analysis of the LBP-ELISA and vaccine protection rate revealed that neutralizing Ab titers greater than 256 provided 100% protection, while titers of 64 and 128 offered 57.1% and 70% protection, respectively. The nbLPB-ELISA is a rapid, simple, safe, and cost-effective method of detecting SVA as a replacement for the SVA VNT and vaccine-challenge assays.

• We evaluated a specific, high-affinity, and neutralizing nanobody-targeting SVA.

• The developed nbLPB-ELISA can replace viral neutralization tests.

• The nbLPB-ELISA is suitable to evaluate protective immunity.

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基于中和纳米抗体的液相阻断酶联免疫吸附试验评估塞内卡病毒 A 疫苗的保护效力

塞内卡病毒A （SVA）引起猪蹄冠水疱病、流产综合征和仔猪死亡率增加。接种疫苗是预防SVA感染的最佳手段。然而，目前对SVA疫苗有效性的评估依赖于用SVA挑战猪，这是耗时、劳动密集型的，并且与动物福利法规不一致。为了替代病毒中和试验（VNT）和SVA攻毒试验，我们建立了以V1-VHH为包被抗体（Ab），生物素化V1-VHH为检测抗体的液相阻断酶联免疫吸附试验（nbLPB-ELISA）。在优化条件下，VNT和nbLPB-ELISA结果呈强相关(Pearson R2 = 0.84；p < 0.00001)。LBP-ELISA分析和疫苗保护率分析显示，中和抗体滴度大于256的保护率为100%，而滴度为64和128的保护率分别为57.1%和70%。nbLPB-ELISA是一种快速、简单、安全且具有成本效益的SVA检测方法，可替代SVA VNT和疫苗激发试验。•我们评估了一种特异性的、高亲和力的、中和的纳米体靶向SVA。•开发的nbLPB-ELISA可替代病毒中和试验。•nbLPB-ELISA适用于保护性免疫评价。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Applied Microbiology and Biotechnology 工程技术-生物工程与应用微生物

CiteScore

10.00

自引率

4.00%

发文量

535

审稿时长

2 months

期刊介绍： Applied Microbiology and Biotechnology focusses on prokaryotic or eukaryotic cells, relevant enzymes and proteins; applied genetics and molecular biotechnology; genomics and proteomics; applied microbial and cell physiology; environmental biotechnology; process and products and more. The journal welcomes full-length papers and mini-reviews of new and emerging products, processes and technologies.