College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, China; Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China.
Henan Provincial Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China.
Int J Biol Macromol. 2023 Dec 31;253(Pt 6):127276. doi: 10.1016/j.ijbiomac.2023.127276. Epub 2023 Oct 5.
Porcine Epidemic diarrhea virus (PEDV), which can result in severe vomiting, diarrhea, dehydration and death in newborn piglets, poses a great threat to the pig industry around the world. The S1 subunit of S protein is crucial for triggering neutralizing antibodies binding to the receptor. Based on the advantages of high immunogenicity and precise assembly of nanoparticles, the mi3 nanoparticles and truncated S1 protein were assembled by the SpyTag/SpyCatcher system and then expressed in HEK293F cells, whereafter high-efficiency monoclonal antibodies (mAbs) were produced and identified. The obtained five mAbs can bind to various genotypes of PEDV, including a mAb (12G) which can neutralize G1 and G2 genotypes of PEDV in vitro. By further identification of monoclonal antibody target sequences, FNDHSF and LFYNVTNSYG were first identified as B-cell linear epitopes, in which LFYNVTNSYG was a neutralizing epitope. Alanine scans identified the key amino acid sites of two epitopes. Moreover, the results of multiple sequence alignment analysis showed that these two epitopes were highly conserved in various subtype variants. In brief, these findings can serve as a basis for additional research of PEDV and prospective resources for the creation of later detection and diagnostic techniques.
猪流行性腹泻病毒（PEDV）可导致新生仔猪严重呕吐、腹泻、脱水和死亡，对全球养猪业构成巨大威胁。S 蛋白的 S1 亚单位对于触发与受体结合的中和抗体至关重要。基于纳米颗粒高免疫原性和精确组装的优势，mi3 纳米颗粒和截短的 S1 蛋白通过 SpyTag/SpyCatcher 系统组装，然后在 HEK293F 细胞中表达，随后产生并鉴定了高效单克隆抗体（mAbs）。获得的 5 种 mAbs 可与包括PEDV G1 和 G2 基因型在内的各种基因型的 PEDV 结合，其中一种 mAb（12G）可在体外中和 PEDV G1 和 G2 基因型。通过进一步鉴定单克隆抗体的靶序列，首次鉴定出 FNDHSF 和 LFYNVTNSYG 为 B 细胞线性表位，其中 LFYNVTNSYG 为中和表位。丙氨酸扫描确定了两个表位的关键氨基酸位点。此外，多序列比对分析的结果表明，这两个表位在各种亚型变体中高度保守。总之，这些发现可以为 PEDV 的进一步研究提供依据，并为后期检测和诊断技术的创建提供潜在资源。
