Protein expression in insect cells

昆虫细胞蛋白表达

强耀生物科技有限公司已成功构建完整的各种蛋白表达技术平台:包括原核蛋白表达与纯化、酵母蛋白表达与纯化、昆虫细胞蛋白表达与纯化、哺乳动物细胞蛋白表达与纯化。根据客户的需求,我们提供从基因合成,载体构建,基因表达,蛋白纯化、检测等一站式服务。为了保证蛋白纯化的质量,强耀生物建立了严格的产品质控体系及一整套完善的客户服务流程、保密制度以确保为客户提供优质的产品及成熟的服务。 我们的蛋白质团队在优化基因密码子、表达载体的设计和构建、融合对象和标签的选取、特定蛋白纯化系统的选择、蛋白生产工艺的优化等方面,拥有较多的经验。可以满足客户个性化的需求。

昆虫细胞蛋白表达与纯化

昆虫细胞表达系统是应用较多的真核表达系统,它可以在昆虫细胞中表达多种外源基因,包括真菌,植物,细菌,病毒的基因。杆状病毒是一类以昆虫细胞为天然宿主的双链DNA病毒,具有高度的种属特异性,不感染脊椎动物,对人、畜无害。目前研究较多的是苜蓿银纹夜蛾核多角体 (AcNPV) ,其宿主是草地贪夜蛾。而sf9细胞,正是杆状病毒表达系统的宿主细胞之一。sf9细胞属于半贴壁型细胞系,可以进行悬浮培养,也可以进行贴壁培养,在应用于蛋白的表达与制备纯化中,有着许多的优点。

1、与其他真核表达系统相比,外源基因表达水平高,特别是细胞内蛋白。多数情况下,表达的重组蛋白可溶,折叠正确有翻译后修饰(如糖基化等),有生物学活性,容易分离纯化;
2、昆虫细胞悬浮生长,容易放大培养,有利于大规模表达重组蛋白;
3、易于表达异源多聚体蛋白,可通过多种重组病毒同时感染昆虫细胞或用含有多个表达框的一种病毒感染昆虫细胞来实现;
4、安全性。昆虫杆状病毒不感染脊椎动物。

强耀生物科技有限公司提供杆状病毒表达载体构建、表达检测以及从大容量细胞培养表达和蛋白纯化等一站式蛋白表达技术服务。

1、克隆目标蛋白编码序列到昆虫杆状病毒转移载体上;
2、制备重组质粒;
3、转染昆虫细胞,制备病毒;
4、病毒滴度测定;
5、重组蛋白表达验证和条件优化;
6、小规模蛋白表达和纯化(500 ml培养物);
7、大规模蛋白表达和纯化(可选,适合于生物医药研究企业)。
相关技术服务:
昆虫杆状病毒制备技术服务
蛋白表达优化技术服务
1、由客户提供5-10ml重组杆状病毒;
2、感染昆虫细胞获得高滴度病毒;
3、表达条件优化。
优化因素包括:
不同的昆虫细胞系;
不同的表达时间;
不同的感染复数(MOI);
病毒滴度测定服务;
空斑实验测定滴度。

Protein expression in insect cells

Protein expression in insect cells

Chinapeptides has successfully built a complete variety of protein expression technology platform, including prokaryotic, yeasty, entomic and mammalian protein expression and purification According to customer needs, we offer gene synthesis, vector, gene expression, protein purification, detection and other one-stop service. In order to ensure the quality of protein purification, Chinapeptides establish a strict quality control system and a set of comprehensive customer service processe, to ensure the quality products and excellent service. Our team has a very rich experience in protein gene codon optimization, vector construction design and expression, integration and label, specific protein purification systems optimization, to meet the needs of individual customers.

Insect Cell Protein Expression and Purification

Insect cell expression system is widely-used eukaryotic expression system that can express a variety of exogenous genes including fungi, plants, bacteria, virus genes. Baculovirus is a type of double-stranded DNA that make insect cells as the natural host. Baculovirus is highly species-specific, and do not infect vertebrate, human, animal. More current research are about Autographa californica nuclear polyhedrosis (AcNPV), which host Spodoptera frugiperda. sf9 cells, one of the host cell for baculovirus expression system. sf9 cells, kind of semi-adherent cell line, can be suspension culture or adherent culture with many many advantages。

1、 compared with other eukaryotic expression systems, it has high exogenous gene expression level, especially intracellular proteins. In cases,recombinant protein is soluble, correctly folding have posttranslational modifications (such as glycosylation, etc.), biologically activity, easy separation and purification;
2、insect cells grown in suspension, which is easy for scaled-up cultures and large-scale expression of recombinant protein;
3、it is easy to express heterologous multimeric proteins. Insect cells can be infected through a variety of recombinant virus or can be implemented with a virus containing a plurality of expression cassettes;
4、security. Baculovirus infection do not include vertebrates.

Chinapeptides provides baculovirus expression vector, expression detecting and mass expression and purification of protein and other one-stop technical services.

1、coding sequence of target protein was cloned into the baculovirus transfer vector;
2、Preparation of a recombinant plasmid
3、transfected into insect cells, the virus modification;
4、viral titer test;
5、the recombinant protein expression verification and Optimization;
6、small-scale protein expression and purification (500 ml culture);
7、large-scale protein expression and purification (optional, suitable for biomedical research enterprise).
Technical services:
Baculovirus technical services
Protein expression optimization services
1、5-10ml recombinant baculovirus is provided by the customer;
2、insect cells are infected to obtain high titer virus;
3、optimization of expression conditions.
Optimization factors include:
Different insect cell lines;
Different expression time;
Different multiplicity of infection (MOI);
Virus titer test services;
Plaque assay titer test.
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