Aarthi Yesupatham, Anjana Aravindha, Tejal Glaudia, Shanmugaraja Meenakshi, Ramadevi S, Princess R
Faculty of Allied Health Sciences, Chettinad Hospital and Research Institute, Chettinad Academy of Research and Education, Kelambakkam, Chennai, Tamil Nadu, 603103, India.
Department of Biotechnology, Mepco Schlenk Engineering College (Autonomous), Sivakasi, Tamil Nadu, 626005, India.
Mol Biotechnol. 2024 Aug 20. doi: 10.1007/s12033-024-01260-0.
A bacterial ghost cell is an empty cell envelope of bacteria lacking cytoplasmic content. Bacterial ghost cells (BGs) can be used for various applications such as vaccines, adjuvants, and drug delivery systems. Since BGs offer many advantages over classically prepared vaccines, developing novel methods for the preparation of high-quality BGs remains to be an interesting field of study by various research groups. Several novel methodologies have been reported that involve the biological (gene E mediated) and combination of various chemicals such as NaOH, SDS, HO, CaCO, and ethanol, non-detergent method using Tween80, limulus antimicrobial peptide, and high hydrostatic pressure method, the porcine myeloid antimicrobial peptide (PMPA) 36-lysozyme fusion method, NaOH-Penicillin/Streptolysin combination method. In this study, we have reported a novel methodology that combines the action of chemical and physical factors to produce ghost cells from gram-negative bacteria, the probiotic E.coli Nissle 1917. The mild detergent Triton X-100 and NaCl alter the permeability of the cell membrane which is further amplified by heat shock induction. This enables the cell to expel its cytoplasmic components without affecting the external morphology. The efficiency of this method was analyzed based on viability assay, cell leakage assay, live-dead cell assay, and scanning electron microscopic analysis. Moreover, the protein loading capacity was optimized for Mycobacterium tuberculosis antigen namely, ESAT-6.
细菌“幽灵”细胞是一种缺乏细胞质内容物的细菌空细胞包膜。细菌“幽灵”细胞（BGs）可用于多种应用，如疫苗、佐剂和药物递送系统。由于BGs比传统制备的疫苗具有许多优势，因此开发制备高质量BGs的新方法仍然是各个研究小组感兴趣的研究领域。已经报道了几种新方法，包括生物学方法（基因E介导）以及各种化学物质的组合，如NaOH、SDS、HO、CaCO和乙醇、使用吐温80的非洗涤剂方法、鲎抗菌肽和高静水压方法、猪髓样抗菌肽（PMPA）36 - 溶菌酶融合方法、NaOH - 青霉素/链球菌溶血素组合方法。在本研究中，我们报道了一种新方法，该方法结合化学和物理因素的作用，从革兰氏阴性细菌、益生菌大肠杆菌Nissle 1917产生“幽灵”细胞。温和的洗涤剂Triton X - 100和NaCl改变细胞膜的通透性，热休克诱导进一步放大这种作用。这使得细胞能够排出其细胞质成分而不影响外部形态。基于活力测定、细胞泄漏测定、活死细胞测定和扫描电子显微镜分析对该方法的效率进行了分析。此外，针对结核分枝杆菌抗原ESAT - 6优化了蛋白质负载能力。
