重组蛋白信息 | Thermo Fisher Scientific

What is a recombinant protein?

A recombinant protein is a manufactured protein that has been produced using cells engineered to carry a gene of interest. Recombinant proteins can have the same amino acid sequence as the corresponding naturally occurring (wild-type) protein, or have their sequence modified for improved properties such as solubility or production yield.

What are recombinant proteins used for?

There are myriad applications for recombinants proteins in biomedical research, as well as diagnostics and therapeutics. Recombinant proteins have become an important tool in various areas of science. Laboratory uses for recombinant proteins include:

Cell culture supplements, including growth factors for primary cell expansion, differentiation, or stem cell reprogramming
Generation of organoids and other 3D cell cultures for disease modeling and drug discovery
Expansion of CAR-T cells by stimulation with recombinant cytokines
Investigational tools for probing cellular function and cell signaling pathways
Antigens for immunization and production of antibodies
Components of functional cell-based assays such as cell migration and angiogenesis

Biologically active recombinant proteins

The above applications require that the recombinant protein be biologically active; that is, the protein is recognized by cells and cause changes in those cells. Bioactivity can be measured in an appropriate cell-based assay and quantified as a specific activity or EC₅₀.

Look up international units for PeproTech proteins
Search catalog of bioactive recombinant proteins

Non-bioactive recombinant proteins may be used as standards in protein analysis studies including immunoassays such as western blotting, ELISA, gel shift, and immunohistochemistry.

Types of recombinant proteins

Many biologically important classes of recombinant proteins are available commercially, obviating the need to express, purify, and validate commonly used bioactive recombinant protein(s).

Major types of recombinant proteins include:

Growth factors: a group of intercellular signaling proteins that cells produce and use to communicate. They stimulate a variety of cellular processes including cell division, differentiation, migration and survival. Prominent growth factor families include:
- FGF: fibroblast growth factors
  - Learn more about the FGF family
  - Explore FGF signaling pathways
- VEGF: vascular endothelial growth factors
  - Learn more about VEGF
  - Explore the VEGF signaling pathway
- Neurotrophins: a subset of growth factors that act primarily on cells involved in the brain and nervous system
Cytokines: including interferons, interleukins, pro-inflammatory cytokines, and chemokines critical to immune cell function and communication
Enzymes: bioactive proteins that can perform chemical transformations; includes recombinant proteases, kinases, and nucleases

Learn more about chemokine structure and function
Explore cytokine and growth factor pathways

Recombinant protein manufacturing categories

当为特定应用选择重组蛋白时，了解需要哪种等级的质量控制和文件是有帮助的。

仅供研究使用 (RUO)

研究级重组蛋白适用于多种生命科学研究应用，包括细胞模型开发、维持细胞表型和体外分析。

参见 Gibco PeproTech RUO 蛋白规范

无动物成分

As an additional safeguard against animal pathogen contamination, animal-free recombinant proteins are made using raw materials that are not of animal origin.

Learn more about PeproTech animal free recombinant proteins

GMP recombinant proteins

GMP-manufactured recombinant proteins undergo extensive testing and are accompanied by documentation to support projects/applications that require this level of rigor.

Learn more about PeproGMP recombinant proteins

FAQs about recombinant proteins

如何测定重组蛋白的生物活性？

生物测定旨在测量给定生长因子或细胞因子的生物活性。在大多数情况下，生物测定是基于细胞的检测，会使用不同的指示细胞，例如原代细胞或细胞系。常用的生物测定包括细胞增殖检测、趋化作用检测、细胞因子生成检测和细胞毒性检测。给定细胞因子的生物活性以 ED₅₀ 表达，它代表诱导 50% 最大反应的细胞因子浓度。

以 ED50 表示的比活度与以单位/mg 表示的比活度之间有何关系？

The specific activity of a biologically active protein can be determined using the following equation:
1 x 10⁶ / ED₅₀ (ng/mL) = specific activity (units/mg)

The ED₅₀ (AKA EC₅₀) can be found on the CoA for the recombinant protein, but we advise determining the ED₅₀ of a given recombinant protein in your functional assay system.

For additional information on ED₅₀ and its relationship with specific activity, please refer to our Tech Tip.

应如何复溶重组蛋白？

首先，对容器进行离心，使粉末聚集在管的底部。我们通常建议将其复溶至 0.1 至 1.0 mg/mL 浓度。大多数蛋白可通过加入无菌蒸馏水复溶。但产品数据表或 CoA 将说明何时需要使用水以外的稀释液。也可在这些文档中找到推荐的溶液、载体蛋白浓度和扩展的储存条件。

应如何储存重组蛋白？

通常，我们建议将冻干重组蛋白储存在 –20°C 下，但可在 4°C 或室温下短期储存。复溶蛋白溶液时，我们建议您制备至少含 10 μL 蛋白溶液的工作等份试液，并在 –20°C 至 –80°C 下储存。不允许进行多次反复冻融。有关储存冻干和复溶重组蛋白的产品特定说明，请见产品手册、数据表或检验证书 (CoA)。

为什么我无法在冻干重组蛋白瓶中看到蛋白沉淀？

重组蛋白通常不含载体蛋白或添加剂（如 BSA、HSA、蔗糖等）。因此在冻干过程中，蛋白成品可能为薄薄的，有时肉眼不可见的薄膜形式沉积在小瓶上，而非颗粒状。沉淀物的大小（如果有）与样品瓶中的重组蛋白数量没有直接关系。

为确保蛋白产物的完全回收，在打开冻干重组蛋白瓶之前，建议对其进行离心 20–30 秒，以将可能残留在瓶盖或侧面的任何蛋白聚集到瓶底。复溶后，您可以通过在 SDS-PAGE 上运行少量产品来确认是否存在产品蛋白。通常来说，只要在丙烯酰胺凝胶上上样 10 ng 的蛋白，应该就能看到预期大小的蛋白条带。

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