Song Shi-Jian, Diao Hai-Ping, Zhang Sen-Sen, Kang Sumin, Liu Chang, Kim Seonghan, Yun Jieun, Meng Hui-Xin, Moon Hongju, Kim Woe-Yeon, Kim Ki Hean, Yang Mao-Jun, Kim Kwang Soon, Guo Yong-Feng, Hwang Inhwan
Qingdao Municipal Key Laboratory of Plant Molecular Pharming, Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao, China; Beijing Life Science Academy (BLSA), Beijing, China.
Qingdao Municipal Key Laboratory of Plant Molecular Pharming, Tobacco Research Institute, Chinese Academy of Agricultural Sciences, Qingdao, China.
Plant Commun. 2025 Aug 11;6(8):101418. doi: 10.1016/j.xplc.2025.101418. Epub 2025 Jun 16.
Plant molecular farming is a promising platform for biopharmaceutical production, however, downstream processing remains a challenge due to cost and complexity. In this study, we present natural peptidoglycan nanoparticles (NPNs) derived from Gram-positive lactic acid bacteria as a novel tool for plant-based vaccine purification and delivery. Sequential treatment with trichloroacetic acid and trypsin effectively reduced NPN size, removing residual host subcellular constructs and proteins while preserving protein-binding capacity. Optimizing trimeric protein anchors and trimerization elements for plant-based expression enabled protein binding at low temperatures, minimizing proteolytic degradation. NPNs conjugated with plant-derived hemagglutinin elicited strong humoral immune responses in mice. Additionally, NPNs enhanced the retention of GFP at the injection site and supported efficient polyclonal antibody generation. These findings establish NPNs as a versatile platform for plant-based recombinant vaccine purification and delivery.
植物分子农业是生物制药生产的一个有前景的平台，然而，由于成本和复杂性，下游加工仍然是一个挑战。在本研究中，我们展示了源自革兰氏阳性乳酸菌的天然肽聚糖纳米颗粒（NPNs），作为一种用于植物疫苗纯化和递送的新型工具。用三氯乙酸和胰蛋白酶进行顺序处理可有效减小NPNs的尺寸，去除残留的宿主亚细胞结构和蛋白质，同时保留蛋白质结合能力。优化用于植物表达的三聚体蛋白锚定物和三聚化元件可使蛋白质在低温下结合，最大限度地减少蛋白水解降解。与植物来源的血凝素偶联的NPNs在小鼠中引发了强烈的体液免疫反应。此外，NPNs增强了绿色荧光蛋白在注射部位的保留，并支持高效的多克隆抗体生成。这些发现确立了NPNs作为植物重组疫苗纯化和递送的通用平台。
