Expi293 Expression System

Trust the industry standard for rapid, high-yield transient HEK 293 expression

Every minute counts when it comes to expressing drug candidates. That’s why the Gibco Expi293 Expression System is the tried-and-true choice for transient expression in 293 cells. This rapid, high-yield system allows you to access recombinant 293–derived proteins in just five to seven days, so you get what you need faster.

高密度哺乳动物瞬时蛋白表达

  • 更高的转染效率
  • 高至10X的蛋白得率
  • 增强的细胞生存能力
  • 更小的培养体积

See expression data Free webinar


Expi293™ Expression System testimonial

“I can produce more protein in less volume and less time, that’s fantastic! It’s like enjoying the benefits of a stable cell line system, without going through the hassle of establishing one. Now we scale up and we can get what we need. It’s robust, it’s scalable, and it’s reproducible. It’s great.”  Joe Sheffer, Senior Research Associate at AnaptysBio, Inc.

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Expi293™ 表达系统视频

瞬时蛋白表达——Expi293™表达系统

如何在实验室中构建哺乳动物瞬时蛋白表达实验

Expi293™表达系统 – 近距离接触发明者系列


What’s in the system?

The Expi293 Expression System brings together a high-expressing 293 cell line, a chemically defined serum–free culture medium, and a high-efficiency transfection reagent with specialized enhancers.

  • 2 vials of frozen Gibco Expi293F Cells
  • 1 L of Gibco Expi293 Expression Medium
  • 1 Gibco ExpiFectamine 293 Transfection Kit, sufficient to transfect 1 L of culture
  • Gibco Opti-MEM I Reduced Serum Medium
  • An antibody-expressing positive control vector
     

Get more protein while reducing hands-on time and waste

The Expi293 Expression System is designed to deliver up to 6x more protein production in just one week, compared with other transient 293 systems that can take two weeks or more. And it requires less plasticware, which means less waste and more incubator space.

“In all my years working with transient expression systems the Expi293 Expression System is the first one to achieve 2.3 grams per liter, beating every other HEK293 transient expression system.”  Large pharmaceutical company

Figure 1. Expression of human IgG and Fc-tagged Cripto achieve expression levels of over 1 g/L in the Expi293 Expression System

See expression data

Expi293 MembranePro Expression System

For researchers studying G protein–coupled receptors (GPCRs) and other membrane proteins, the Expi293 MembranePro Expression System is designed to deliver enriched, functional membrane proteins with high efficiency, scalability, and reliability.

  • Efficient—20x greater membrane protein yield compared to the adherent system
  • Scalable—easy scale-up without the need for multiple flasks
  • Easy to use—no enzymatic or mechanical dissociation required

Discover more

Study correctly folded, active protein and assembled complexes

The Expi293 Expression System is based on expression in mammalian 293 cells, which offers the benefits of native folding and posttranslational modifications. Figure 2 shows purification of the HDAC3/SMRT-DAD complex from 293 cells and E. coli. The 293 cells yield a stable pure HDAC3 complex (Figure 2A), whereas expression in E. coli (Figure 2B) does not.

Figure 2. Purification of the HDAC3/SMRT-DAD complex from (A) mammalian 293 cells and (B) E. coli. In both cases the protein complex was purified using an affinity tag on SMRT: Flag-tag for the mammalian-expressed proteins, and GST-tag for the E. coli–expressed proteins. The HDAC3 was untagged.
Equivalent rabbit IgG titers are obtained in cGMP-banked Expi293F cells and catalog Expi293F cells at 30 mL scale using the Expi293 Expression System.

Fully documented cGMP-banked cells

  • Fully documented cGMP-banked cell line 
  • Derived from the same Master Seed Bank as standard Expi293F Cells (Cat No. A14527
  • Pre-adapted to Expi293 Expression Medium 
  • A component of the fully integrated, animal origin-free Expi293 Expression System for applications requiring cGMP-banked cells 
  • Flexible commercial licensing options 

Click here for product information

Get more protein while reducing hands-on time and waste

The Expi293 Expression System is designed to deliver up to 6x more protein production in just one week, compared with other transient 293 systems that can take two weeks or more. And it requires less plasticware, which means less waste and more incubator space.

“In all my years working with transient expression systems the Expi293 Expression System is the first one to achieve 2.3 grams per liter, beating every other HEK293 transient expression system.”  Large pharmaceutical company

Figure 1. Expression of human IgG and Fc-tagged Cripto achieve expression levels of over 1 g/L in the Expi293 Expression System

See expression data

Expi293 MembranePro Expression System

For researchers studying G protein–coupled receptors (GPCRs) and other membrane proteins, the Expi293 MembranePro Expression System is designed to deliver enriched, functional membrane proteins with high efficiency, scalability, and reliability.

  • Efficient—20x greater membrane protein yield compared to the adherent system
  • Scalable—easy scale-up without the need for multiple flasks
  • Easy to use—no enzymatic or mechanical dissociation required

Discover more

Study correctly folded, active protein and assembled complexes

The Expi293 Expression System is based on expression in mammalian 293 cells, which offers the benefits of native folding and posttranslational modifications. Figure 2 shows purification of the HDAC3/SMRT-DAD complex from 293 cells and E. coli. The 293 cells yield a stable pure HDAC3 complex (Figure 2A), whereas expression in E. coli (Figure 2B) does not.

Figure 2. Purification of the HDAC3/SMRT-DAD complex from (A) mammalian 293 cells and (B) E. coli. In both cases the protein complex was purified using an affinity tag on SMRT: Flag-tag for the mammalian-expressed proteins, and GST-tag for the E. coli–expressed proteins. The HDAC3 was untagged.
Equivalent rabbit IgG titers are obtained in cGMP-banked Expi293F cells and catalog Expi293F cells at 30 mL scale using the Expi293 Expression System.

Fully documented cGMP-banked cells

  • Fully documented cGMP-banked cell line 
  • Derived from the same Master Seed Bank as standard Expi293F Cells (Cat No. A14527
  • Pre-adapted to Expi293 Expression Medium 
  • A component of the fully integrated, animal origin-free Expi293 Expression System for applications requiring cGMP-banked cells 
  • Flexible commercial licensing options 

Click here for product information

Use your preferred culture format without sacrificing yield

The Expi293 Expression System is designed to support robust protein production in a variety of culture vessels. In small and large-scale formats—and everything in between—you can express the protein you need.

Figure 3. Expression of Fc-tagged Cripto achieves similar volumetric protein yields at 3 different scales.

 

Table 1. Expi293 is cost-effective compared to other FreeStyle 293 Medium.

NOTE: Cost comparison based on list price in USD and protein yields of 1 g/L with Expi293 system.


Expi293™表达系统应用

高出20倍的膜蛋白得率

Expi293 MembranePro表达系统
对于研究G蛋白偶联受体(GPCR)和其他膜蛋白的研究人员而言,Expi293™ MembranePro™表达系统可以制备出富集的功能性膜蛋白,具有高效率、高可扩展性及高可靠性优势。

  • 高效— 相对于贴壁培养体系,蛋白得率可提高20倍
  • 可扩展—无需采用多培养皿,即可轻松扩大化规模
  • 易于使用—无需酶促或机械解离步骤

了解更多


Precise protein expression is all in the fold

For structural biologists, insoluble and misfolded proteins mean starting over. The Expi293 Expression System is precisely designed with suspension 293 cells, optimized medium and transfection reagent to provide native folding and maximum protein yields. Now, your structural study can get the biological relevance it deserves.

Discover more

The Expi293™ Expression System is a rapid method of producing mg to gram quantities of recombinant protein in mammalian cells. For the expression of human or other mammalian proteins, expression in the human-derived 293 cells offers the benefit of more native folding and post-translational modifications, such as glycosylation, when compared to expression systems based on hosts such as E. coli, yeast, or insect cells.


Expi293™ 实验方案

Expi293™ 表达系统提供了你的实验室轻松制备高水平重组蛋白质的所有所需组件。唯一需要额外提供的组件时含有你感兴趣蛋白质对应DNA的哺乳动物表达载体。

将Expi293F™细胞融解后在Expi293™表达培养基中进行悬浮培养。细胞通常可培养于摇床上的摇瓶中CO2细胞培养保温箱中或生物反应器中。使用ExpiFectamine™ 293转染试剂进行细胞转染。在转染后16-18小时后将转染增强子1及2加入至培养基中。在蛋白制备过程中无需改变培养基或添加培养基。在转染后2至7天内可开始收集蛋白,具体时间取决于感兴趣蛋白的表达方式。对于重组抗体表达来说,常常在转染后5至7天时达到最大得率。

该系统具有高度扩展性,从24孔平板中的1mL培养体系到摇瓶中的1L培养基中均可得到相似的蛋白得率(参见图1)。

Expression of Fc-tagged Cripto  
图1. 3种不同培养规模下Fc标记Crip蛋白的表达可达到相似的蛋白得率。


应用说明

高出20倍的膜蛋白得率

Expi293 MembranePro表达系统
对于研究G蛋白偶联受体(GPCR)和其他膜蛋白的研究人员而言,Expi293™ MembranePro™表达系统可以制备出富集的功能性膜蛋白,具有高效率、高可扩展性及高可靠性优势。

  • 高效— 相对于贴壁培养体系,蛋白得率可提高20倍
  • 可扩展—无需采用多培养皿,即可轻松扩大化规模
  • 易于使用—无需酶促或机械解离步骤

了解更多


Precise protein expression is all in the fold

For structural biologists, insoluble and misfolded proteins mean starting over. The Expi293 Expression System is precisely designed with suspension 293 cells, optimized medium and transfection reagent to provide native folding and maximum protein yields. Now, your structural study can get the biological relevance it deserves.

Discover more

The Expi293™ Expression System is a rapid method of producing mg to gram quantities of recombinant protein in mammalian cells. For the expression of human or other mammalian proteins, expression in the human-derived 293 cells offers the benefit of more native folding and post-translational modifications, such as glycosylation, when compared to expression systems based on hosts such as E. coli, yeast, or insect cells.


Expi293™ 实验方案

Expi293™ 表达系统提供了你的实验室轻松制备高水平重组蛋白质的所有所需组件。唯一需要额外提供的组件时含有你感兴趣蛋白质对应DNA的哺乳动物表达载体。

将Expi293F™细胞融解后在Expi293™表达培养基中进行悬浮培养。细胞通常可培养于摇床上的摇瓶中CO2细胞培养保温箱中或生物反应器中。使用ExpiFectamine™ 293转染试剂进行细胞转染。在转染后16-18小时后将转染增强子1及2加入至培养基中。在蛋白制备过程中无需改变培养基或添加培养基。在转染后2至7天内可开始收集蛋白,具体时间取决于感兴趣蛋白的表达方式。对于重组抗体表达来说,常常在转染后5至7天时达到最大得率。

该系统具有高度扩展性,从24孔平板中的1mL培养体系到摇瓶中的1L培养基中均可得到相似的蛋白得率(参见图1)。

Expression of Fc-tagged Cripto  
图1. 3种不同培养规模下Fc标记Crip蛋白的表达可达到相似的蛋白得率。


应用说明


Expi293™ 表达系统构成

Expi293™ Expression System

ExpiFectamine™ 293 试剂及ExpiFectamine™ 293转染增强子1和2以ExpiFectamine™ 293转染试剂盒的形式一同销售。其他组分均可单独购买。该试剂盒中包含:

  • 2瓶冷冻的Expi293F™细胞
  • 1升Expi293™表达培养基
  • 1盒ExpiFectamine™ 293转染试剂盒,足以转染1升培养细胞
  • Opti-MEM®低血清培养基
  • 抗体表达阳性对照

Expi293™ 表达系统 是瞬时表达技术的一大进步,可在哺乳动物细胞中进行快速的超高得率蛋白制备。该系统基于Expi293F™细胞在Expi293™表达培养基上的高密度培养。瞬时表达则由全新的阳离子脂质体ExpiFectamine™ 293转染试剂与经优化的转染增强子的结合使用来驱动,这种增强子专门设计用于与该款转染试剂协同使用。所有这些成分协调作用使得蛋白得率相比常规表达系统,如FreeStyle™ 293系统等,可高出2至10倍。进行IgG及非IgG蛋白表达时表达水平可超过1 g/L(参见图2)。
 

Expi293™ Expression System  
图2. 人IgG和Fc标记的Crip蛋白在Expi293™表达系统中的表达水平均超过了1 g/L。


Expi293™表达系统的特性

Expi293™ 表达培养基是一种化学成分明确的无血清无蛋白培养基。Expi293™表达培养基可用于高密度细胞培养(参见图3)和转染,相比传统培养系统可达到更高的蛋白得率。

Expi293™ Expression Medium  
图3. 生长于Expi293™表达培养基与其他293培养基的细胞随时间细胞生存能力及存活细胞密度的变化。

Expi293F™ 细胞经特别适应性改造,相比传统293细胞可表现出更高的生产能力(pg/cell/day),在用于Expi293™表达系统时相比Freestyle™ 293F细胞可制得1.7x的蛋白质。ExpiFectamine™ 293 转染试剂适用于高密度293培养体系的高效转染,可配合转染增强子适用来进一步提高转染效果和表达水平(参见图4)。
 

Expression of hIgG  
图4. 使用或不使用转染增强子情况下的hIgG表达。

Thermo Scientific Nalgene sterile single-use Erlenmeyer flasks

Sterile, single-use Thermo Scientific Nalgene flasks are made from light, break-resistant, crystal-clear PETG, so you can express with confidence. All sizes are available in vented or non-vented formats, and with plain or baffled bottoms for use on the benchtop or shaker table. See protocol for flasks validated with Expi293 Expression system.

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