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Recombinant proteins and expression systems

Selecting the most appropriate expression system is important when choosing a recombinant protein. The wrong choice can lead to inactive proteins that do not function correctly in downstream

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Explore our extensive range of high-quality proteins, designed to give you consistent and accurate results for a variety of applications. 

Find your bioactive protein

Last edited Thu 01 Sep 2022

What are recombinant bioactive proteins, and how are they produced?

Recombinant proteins are manipulated versions of native proteins produced using recombinant DNA to generate large amounts of a specific protein. The recombinant DNA is cloned into a vector introduced in a specific expression system (eg, mammalian, bacteria, yeast, or insect cells) to support the expression of the gene of interest and the production of the recombinant protein. Specifically, a bioactive protein is a protein that elicits an effect or response in a biological system.

Pure, high-quality proteins with high batch-to-batch consistency are critical for reliable results in biological research. When purchasing a commercially produced recombinant protein, it is also important to consider the expression system used for production. Each protein expression system has different features and applications and is chosen based on the protein type, function, and required yield.

Abcam offers two ranges of bioactive proteins, depending on your needs:

Bioactive grade proteins: ideal for cell culture and functional studies Premium grade bioactive proteins: ultra-high purity cytokines and growth factors perfect for preclinical cell culture and functional studies 
  • Validated bioactivity
  • Mammalian, E.coli, insect, yeast, or wheat germ cell-free expression system
  • High purity (>90%) confirmed by SDS-PAGE
  • Endotoxin level below <1 EU per µg
  • Size confirmed by SDS-PAGE
  • Optimal bioactivity
  • Ultra-high purity confirmed by LC/MS (>95%) and SDS-PAGE (>99%)
  • Ultra-low endotoxin (<0.005 EU per µg)
  • Size confirmed by mass spectrometry
  • Animal-free
  • Carrier free
  • Tags removed
  • Manufactured to ISO 9001:2015

We have an extensive range of high-quality proteins made in various expression systems, so you can find a protein that fits your experimental needs.

Which expression system is best for your needs?

There are many expression systems across the biological kingdoms, including cell-free methods, and systems using cell lines from bacteria, plants, fungi, insects, and mammals (including humans).

Proteins produced in prokaryotes do not undergo post-translational modifications (PTMs) such as glycosylation, phosphorylation, or proteolytic processing. PTMs are involved in folding processes, stability, and biological activity. Although prokaryotic expression systems are suitable for many applications, human-origin proteins produced in bacterial systems may not be folded or modified correctly, so they won’t necessarily function in the same way as their native-identical counterparts nor be as stable.

Prokaryotic-produced proteins are less expensive to manufacture but could be more costly over the lifecycle of a project. The expression system used when proteins are manufactured can impact activity, usefulness in downstream applications, and protein stability. Understanding these factors are crucial to ensure you don’t experience extra expense when purchasing proteins. It may be more cost-effective to start with a more costly protein that will save you money in the long run due to its increased stability (and therefore shelf life) or higher levels of activity.  

Below we introduce the most popular protein expression systems for recombinant protein production, with a brief summary of the key features of each method.  

Expression systemUsesAdvantagesLimitations
Mammalian
  • Assays requiring functional or larger, more complex proteins, eg, functional and structural assays, protein interaction studies
  • Antibody generation
  • Preclinical cell culture
  • High level processing, eg, PTMs and correct folding
  • Produces bioactive, stable proteins effectively identical to the human native version with ultra-low endotoxin levels
  • Optimal bioactivity
  • High purity
  • More complex culture conditions
  • High yields are only possible in suspension culture
Insect
  • Assays requiring functional or larger, more complex proteins, eg, functional and structural assays, protein interaction studies
  • Expressing intracellular proteins and protein complexes
  • Vaccine and diagnostic assay development
  • Can use relatively high cell densities in culture, giving higher yields
  • Processing similar to mammalian proteins
  • More difficult to culture than bacteria or yeast
  • Production of recombinant baculovirus can be time consuming
E. coli
  • Structural or functional analysis for small, low-cost projects
  • Protein interaction studies
  • Antibody generation
  • Screening applications
  • Easy to culture
  • Production is rapid and scalable
  • Eukaryotic proteins may be incorrectly folded and form insoluble inclusion bodies
  • Mammalian PTMs not present in prokaryotic proteins
Yeast
  • Functional and structural analyses
  • Protein interaction studies
  • Antibody generation
  • Eukaryotic PTMs and modifications possible
  • Large scale fermentation possible
  • Some PTMs, eg, glycosylation, are less complex than in mammalian cells
  • Culture may require optimization
Cell free
  • Production of toxic proteins that would kill host cells
  • Proteins incorporating labels
  • Expressing complex proteins such as membrane proteins or those containing disulfide bonds
  • Functional assays
  • Protein interaction studies
  • Screening
  • Rapid production - doesn't require cloning or transfection
  • Can be animal/serum-free (eg, wheatgerm expressed)
  • Large scale production is costly
  • Lack of PTMS and potential incorrect folding