Tissue specific and safety pinpoint targeting by bionano-capusele

介绍：生物纳米胶囊（BNC）是我们通过生物技术程序设计和生产的人造中空纳米颗粒的概念。由于L-蛋白氨基端对人肝细胞具有特异性亲和力，因此只有纳米尺寸的中空颗粒显示出极高的选择性靶向潜力，作为一种新型的直接靶向人肝的DDS载体。由于其良好的潜力，我们提出了BNC概念作为一种有效的纳米机器，实现基因，药物和蛋白质的组织/细胞型特异性delivery.结果：1.通过过表达的L蛋白，这是有效的分泌信号肽的L颗粒。在无抗生素选择压力的无血清培养基中培养的5天内，L颗粒的浓度达到约1.7 g/ml。分泌产生的L颗粒有利于他们容易通过层析纯化。此外，D ...更多信息 证明纯化的L颗粒仅能转染人肝细胞。因此，昆虫细胞表达系统是一种有吸引力的生产BNC的工具。2.连续全身性地用所得到的FGF和RNase的昆虫融合蛋白（CL-RFN 89）治疗体内的人A431鳞状细胞癌显著抑制生长。与对照小鼠相比，用CL-RFN 89处理7天导致58.2%的肿瘤生长抑制（p<0.001）。3.荧光素标记的hRNase 1-hEGF可被EGFR过表达的A431细胞内化。hEGF处理的A431细胞和EGFR缺陷的H69细胞均未观察到内化。4.本研究设计了绿色荧光蛋白（EGFP）与S区C端的融合体。优化C-末端的截短以在Cos 7细胞中获得足够的表达水平。通过免疫反应性、荧光和条件培养基和细胞提取物中的密度来评价所产生的纳米颗粒。我们发现两个方向的EGFP部分在BNC中，是展示EGFP外，并将其包围内，而只有一个方向被发现占据一个粒子，使任何类型的BNC-EGFPs可以有效地和独立地分离抗体亲和column.Conclusion：这种重组BNC正在开发作为一种新的平台的药物输送系统载体的选择性交付。少

Introduction : The bio-nanocapsule (BNC) is our concept of artificial hollow nanoparticles that have been designed and produced through biotechnological procedures. Due to the specific affinity to human hepatocytes localized in the amino-terminus of L-protein, only the nanometer size hollow particles, showed extremely selective targeting potential as a novel type of DDS vector to the directed to the human liver. Because of its favorable potential, we have proposed a BNC concept as an efficient nanomachine to achieve tissue-/cell-type specific delivery of genes, drugs and proteins.Result : 1.L particles were efficiently secreted by the overexpression of the L protein, which was fused to the secretion signal peptide. The concentration of L particles was reached approximately 1.7 g/ml in 5 days during cultivation in a serum-free medium without antibiotic selective pressure. The secretory production of L particles facilitated their easy purification by chromatography. Furthermore, it was d … More emonstrated that purified L particles can transfect only human hepatocytes. Therefore, an insect cell expression system is an attractive tool for the production of BNC.2.Continuous systemic treatment with resultant insetional-fusion protein of FGF and RNase (CL-RFN89) significantly inhibited the growth of human A431 squamous cell carcinomas in vivo. Seven days of treatment with CL-RFN89 resulted in a 58.2% inhibition of tumor growth compared with control mice (p<0.001). Furthermore, immunohistochemistry using a rat anti-mouse CD31 antibody snowed that treatment with CL-RFN89 reduced tumor vascularization.3.FITC-labelled hRNase 1-hEGF was internalized into EGFR-overexpressing A431 cells. The internalization was not observed In A431 cells pre-treated with hEGF and EGFR-deficient H69 cells.4.The fusion of jellyfich green fluorescent protein (EGFP) to the C-terminus of the S region was designed in this study. Truncation of the C-terminus was optimized to obtain sufficient expression level in Cos 7 cells. The produced nanoparticles were evaluated by immunoreactivity, fluorescence, and density both in the conditioned media and in the cell extracts. We found two orientations of the EGFP moiety in BNCs that are displaying EGFP outside and are enclosing it inside, while only single orientation was found to occupy a particle, so that either type of BNC-EGFPs could be effectively and independently separated by antibody affinity column.Conclusion : This recombinant BNC is now being developed as a novel platform of drug delivery system vector for selective delivery. Less