Sabet Salwa, George Marina A, El-Shorbagy Haidan M, Bassiony Heba, Farroh Khaled Y, Youssef Tareq, Salaheldin Taher A
Department of Zoology, Faculty of Science, Cairo University, Giza, Egypt.
Department of Photochemistry and Photobiology, National Institute of Laser Enhanced Sciences, Cairo University, Giza, Egypt.
PLoS One. 2017 Jul 26;12(7):e0181723. doi: 10.1371/journal.pone.0181723. eCollection 2017.
Development of an effective non-viral vaccine against hepatitis C virus infection is of a great importance. Gelatin nanoparticles (Gel.NPs) have an attention and promising approach as a viable carrier for delivery of vaccine, gene, drug and other biomolecules in the body.
The present study aimed to develop stable Gel.NPs conjugated with nonstructural protein 2 (NS2) gene of Hepatitis C Virus genotype 4a (HCV4a) as a safe and an efficient vaccine delivery system.
Gel.NPs were synthesized and characterized (size: 150±2 nm and zeta potential +17.6 mv). NS2 gene was successfully cloned and expressed into E. coli M15 using pQE-30 vector. Antigenicity of the recombinant NS2 protein was confirmed by Western blotting to verify the efficiency of NS2 as a possible vaccine. Then NS2 gene was conjugated to gelatin nanoparticles and a successful conjugation was confirmed by labeling and imaging using Confocal Laser Scanning Microscope (CLSM). Interestingly, the transformation of the conjugated NS2/Gel.NPs complex into E. coli DH5-α was 50% more efficient than transformation with the gene alone. In addition, conjugated NS2/Gel.NPs with ratio 1:100 (w/w) showed higher transformation efficiency into E. coli DH5-α than the other ratios (1:50 and 2:50).
Gel.NPs effectively enhanced the gene delivery in bacterial cells without affecting the structure of NS2 gene and could be used as a safe, easy, rapid, cost-effective and non-viral vaccine delivery system for HCV.
开发一种有效的抗丙型肝炎病毒感染的非病毒疫苗具有重要意义。明胶纳米颗粒(Gel.NPs)作为一种可行的载体,用于在体内递送疫苗、基因、药物和其他生物分子,受到关注且前景广阔。
本研究旨在开发与丙型肝炎病毒4a基因型(HCV4a)的非结构蛋白2(NS2)基因偶联的稳定明胶纳米颗粒,作为一种安全有效的疫苗递送系统。
合成并表征了明胶纳米颗粒(尺寸:150±2 nm,ζ电位 +17.6 mv)。使用pQE - 30载体成功将NS2基因克隆并表达至大肠杆菌M15中。通过蛋白质免疫印迹法确认重组NS2蛋白的抗原性,以验证NS2作为潜在疫苗的有效性。然后将NS2基因与明胶纳米颗粒偶联,并使用共聚焦激光扫描显微镜(CLSM)进行标记和成像,证实偶联成功。有趣的是,偶联的NS2/Gel.NPs复合物转化至大肠杆菌DH5-α的效率比单独基因转化高50%。此外,比例为1:100(w/w)的偶联NS2/Gel.NPs转化至大肠杆菌DH5-α的效率高于其他比例(1:50和2:50)。
明胶纳米颗粒有效地增强了细菌细胞中的基因递送,且不影响NS2基因的结构,可作为一种安全、简便、快速、经济高效的非病毒丙型肝炎疫苗递送系统。