Improving the baculovirus expression vector system with vankyrin-enhanced technology.
Improving the baculovirus expression vector system with vankyrin-enhanced technology.
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通过Vankyrin增强技术改善杆状病毒表达载体系统。
DOI：
10.1002/btpr.2516
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发表时间：
2017-11
影响因子：
2.9
通讯作者：
Geisler C
中科院分区：
文献类型：
--
作者：
Steele KH;Stone BJ;Franklin KM;Fath-Goodin A;Zhang X;Jiang H;Webb BA;Geisler C
杆状病毒表达载体系统（BEVS）是一种广泛用于生产重组真核蛋白的平台。然而，与其他高等真核表达系统相比，BEVS具有局限性。首先，BEVS中使用的昆虫细胞系不能产生具有复杂型N-糖基化模式的糖蛋白。第二，蛋白质的产生是有限的，因为细胞响应于杆状病毒感染而死亡和裂解。为了延迟细胞死亡和裂解，我们用携带vankyrin基因（P-vank-1）的表达质粒转化了几种昆虫细胞系，该基因编码抗凋亡蛋白。具体来说，我们转化了Sf 9细胞、粉纹夜蛾High FiveTM细胞和SfSWT-4细胞，这些细胞可以产生具有复杂型N-糖基化模式的糖蛋白。包括后者的目的是增加具有复杂N-聚糖的糖蛋白的产生，从而克服BEVS的两个上述限制。为了进一步增加vankyrin表达水平并进一步延迟细胞死亡，我们还用P-vank-1基因修饰了杆状病毒载体。我们发现，当SfSWT-4细胞感染编码vankyrin的杆状病毒时，细胞裂解延迟，重组糖蛋白产量增加。当表达vankyrin的细胞与编码vankyrin的杆状病毒组合时，观察到重组蛋白产量水平升高的协同效应。用各种模型蛋白质（包括医学相关的治疗性蛋白质）观察到这些作用。总之，我们发现，通过使用组成型表达vankyrin或编码vankyrin的杆状病毒载体的细胞系，可以延迟细胞裂解并增加重组蛋白产量。© 2017 The Authors Biotechnology Progress出版由Wiley Periodicals，Inc.代表美国化学工程师生物技术协会。程序：33：1496-1507，2017
The baculovirus expression vector system (BEVS) is a widely used platform for the production of recombinant eukaryotic proteins. However, the BEVS has limitations in comparison to other higher eukaryotic expression systems. First, the insect cell lines used in the BEVS cannot produce glycoproteins with complex‐type N‐glycosylation patterns. Second, protein production is limited as cells die and lyse in response to baculovirus infection. To delay cell death and lysis, we transformed several insect cell lines with an expression plasmid harboring a vankyrin gene (P‐vank‐1), which encodes an anti‐apoptotic protein. Specifically, we transformed Sf9 cells, Trichoplusia ni High FiveTM cells, and SfSWT‐4 cells, which can produce glycoproteins with complex‐type N‐glycosylation patterns. The latter was included with the aim to increase production of glycoproteins with complex N‐glycans, thereby overcoming the two aforementioned limitations of the BEVS. To further increase vankyrin expression levels and further delay cell death, we also modified baculovirus vectors with the P‐vank‐1 gene. We found that cell lysis was delayed and recombinant glycoprotein yield increased when SfSWT‐4 cells were infected with a vankyrin‐encoding baculovirus. A synergistic effect in elevated levels of recombinant protein production was observed when vankyrin‐expressing cells were combined with a vankyrin‐encoding baculovirus. These effects were observed with various model proteins including medically relevant therapeutic proteins. In summary, we found that cell lysis could be delayed and recombinant protein yields could be increased by using cell lines constitutively expressing vankyrin or vankyrin‐encoding baculovirus vectors. © 2017 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers Biotechnol. Prog., 33:1496–1507, 2017
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影响因子：
13.8
作者：
Bieniossek C;Imasaki T;Takagi Y;Berger I
通讯作者：
Berger I
影响因子：
2.9
作者：
Contreras-Gomez, A.;Sanchez-Miron, A.;Chisti, Y.
通讯作者：
Chisti, Y.
影响因子：
5.4
作者：
Chen, Yun-Ru;Zhong, Silin;Blissard, Gary W.
通讯作者：
Blissard, Gary W.
DOI：
10.1007/978-1-4939-2760-9_10
发表时间：
2015-01-01
期刊：
GLYCO-ENGINEERING: METHODS AND PROTOCOLS
影响因子：
--
作者：
Geisler, Christoph;Mabashi-Asazuma, Hideaki;Jarvis, Donald L.
通讯作者：
Jarvis, Donald L.
影响因子：
2.6
作者：
Fath-Goodin, A.;Kroemer, J. A.;Webb, B. A.
通讯作者：
Webb, B. A.