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EDTA

0.5M EDTA

Company: Quality Biological
Catalog#: 351-027-721
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Preparation of a Bacteriophage T4-based Prokaryotic-eukaryotic Hybrid Viral Vector for Delivery of Large Cargos of Genes and Proteins into Human Cells
Author:
Date:
2020-04-05
[Abstract]  A viral vector that can safely and efficiently deliver large and diverse molecular cargos into cells is the holy grail of curing many human diseases. Adeno-associated virus (AAV) has been extensively used but has a very small capacity. The prokaryotic virus T4 has a large capacity but lacks natural mechanisms to enter mammalian cells. Here, we created a hybrid vector by combining T4 and AAV into one nanoparticle that possesses the advantages of both. The small 25 nm AAV particles are attached to the large 120 nm x 86 nm T4 head through avidin-biotin cross-bridges using the phage decoration proteins Soc (small outer capsid protein) and Hoc (highly antigenic outer capsid protein). AAV thus “piggy-backed” on T4 capsid, by virtue of its natural ability to enter many types of human cells ... [摘要]  [摘要 ] 一种病毒载体,可以安全有效地将大量多样的分子货物运送到细胞中 是治愈许多人类疾病的圣杯。腺伴随病毒(AAV)已被广泛使用,但容量很小。T4原核病毒容量大,但缺乏进入哺乳动物细胞的天然机制。在这里,我们通过将T4和AAV结合到一个具有两者优势的纳米颗粒中,创建了一种杂交载体。使用噬菌体修饰蛋白Soc(小的外衣壳蛋白)和Hoc(高度抗原化的外衣壳蛋白),通过亲和素-生物素交叉桥将25 nm的AAV小颗粒连接到120 nm x 86 nm的大T4头上。因此,AAV凭借其固有的进入多种类型人体细胞的自然能力,可以“背负”于T4衣壳上,从而有效地充当了“驱动器”,以运送与T4头相关的大型货物。这种独特的T4-AAV杂交载体方法可为将来开发新型疗法铺平道路。

[背景 ] 已经有新的和有效的递送载体能够运输基因和蛋白质的大货物进入人类细胞,以刺激生产治疗性生物分子的和/或修复的细胞和遗传缺陷的迫切需要。这样的载体将允许将快速出现的技术(例如CRISPR,CAR T细胞等)转化为用于大规模应用以及个性化医学的疗法(Stewart 等,2016)。

将具有不同特性的纳米粒子组装到杂化复合物中是开发新型功能材料的有力策略,因为这些杂化复合物显示出集体和协作的属性,其中某些属性可能与单个粒子所显示的属性不同(Ghosh 等人,2012; ...

Isolation of Mouse Tumor-Infiltrating Leukocytes by Percoll Gradient Centrifugation
Author:
Date:
2013-09-05
[Abstract]  A hallmark of cancer-associated inflammation is the infiltration of leukocytes into tumors, which is believed to be recruited by chemokines. Some infiltrating leukocytes such as macrophages often promote tumor growth by producing growth-inducing and angiogenic factors. Here, we described a method of isolating tumor-infiltrating leukocytes with Percoll density gradient, because Percoll possesses a low viscosity, a low osmolarity and no toxicity to cells. Different leukocyte populations are isolated based on their density and collected at the interface between 40% and 80% discontinuous Percoll gradient. [摘要]  癌症相关炎症的标志是白细胞浸润到肿瘤中,据信这是由趋化因子募集的。 一些浸润性白细胞如巨噬细胞通常通过产生生长诱导和血管生成因子来促进肿瘤生长。 在这里,我们描述了一种用Percoll密度梯度分离肿瘤浸润白细胞的方法,因为Percoll具有低粘度,低渗透压和对细胞无毒性。 基于它们的密度分离不同的白细胞群体,并在40%和80%不连续的Percoll梯度之间的界面处收集。

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