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从人血无细胞提取物中快速重组蛋白表达。

Rapid recombinant protein expression in cell-free extracts from human blood.

机构信息

Center for Advanced Sensor Technology (CAST) and Department of Chemical Biochemical and Environmental Engineering (CBEE), University of Maryland Baltimore County, Baltimore, Maryland, USA.

出版信息

Sci Rep. 2018 Jun 22;8(1):9569. doi: 10.1038/s41598-018-27846-8.

Abstract

Several groups have recently reported on the utility of cell-free expression systems to make therapeutic proteins, most of them employing CHO or E. coli cell-free extracts. Here, we propose an alternative that uses human blood derived leukocyte cell extracts for the expression of recombinant proteins. We demonstrate expression of nano luciferase (Nluc), Granulocyte-colony stimulating factor (G-CSF) and Erythropoietin (EPO) in cell-free leukocyte extracts within two hours. Human blood is readily available from donors and blood banks and leukocyte rich fractions are easy to obtain. The method described here demonstrates the ability to rapidly express recombinant proteins from human cell extracts that could provide the research community with a facile technology to make their target protein. Eventually, we envision that any recombinant protein can be produced from patient-supplied leukocytes, which can then be injected back into the patient. This approach could lead to an alternative model for personalized medicines and vaccines.

摘要

最近有几个小组报告了使用无细胞表达系统来制造治疗性蛋白质的效用,其中大多数使用 CHO 或大肠杆菌无细胞提取物。在这里,我们提出了一种替代方案,使用人血液衍生的白细胞细胞提取物来表达重组蛋白。我们证明了纳米荧光素酶(Nluc)、粒细胞集落刺激因子(G-CSF)和促红细胞生成素(EPO)在无细胞白细胞提取物中的表达可在两小时内完成。人血可从供体和血库中轻易获得,富含白细胞的部分也很容易获得。本文所述的方法证明了从人细胞提取物中快速表达重组蛋白的能力,这可为研究界提供一种简便的技术来制造他们的目标蛋白。最终,我们设想可以从患者提供的白细胞中生产任何重组蛋白,然后将其重新注入患者体内。这种方法可以为个性化药物和疫苗提供一种替代模式。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/05ab/6014972/01ac65efb418/41598_2018_27846_Fig1_HTML.jpg

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