SUMO-Tag Protein Expression & Purification
Advanced protein expression solutions for superior yield, solubility, and native protein folding across prokaryotic and eukaryotic systems.
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- Overview
Protein expression and production of recombinant proteins routinely encounter problems, including formation of inclusion bodies, incorrect protein conformation, toxicity to the host cell, and low protein yield. These issues are most often addressed by using an alternative host or genetically fusing the protein of interest (POI) to a carrier protein (fusion tag).
Attachment of the C-terminus of SUMO (Small Ubiquitin-like Modifier) to the N-terminus of a protein of interest can dramatically improve protein solubility, achieve native protein folding, and increase total yield by improving expression and decreasing degradation.
- SUMO-Tag Expression System
Our SUMO-tag expression system maximizes the yield of soluble, functional proteins in E. coli. SUMO functions as both a chaperone and initiator of protein folding, improving the solubility and yield of a POI. DeSUMOylases remove the SUMO tags, releasing your protein with the desired N-terminal amino acid. Both SUMO tags and deSUMOylases contain 6xHis tags allowing efficient subsequent removal, leaving pure protein.
SUMOpro System
SUMOstar Tag
Enhanced Expression
Since all eukaryotic cells contain SUMO and SUMO proteases, the latter of which will cleave SUMO fusion proteins immediately following expression, LifeSensors has engineered SUMOstar, a SUMO tag for use in eukaryotic hosts, which is resistant to endogenous proteases, thus preserving enhanced expression, increased solubility and native folding. SUMOstar fusion proteins can be cleaved only with SUMOstar protease.
To overcome these challenges, we developed the SUMOstar™ tag, an engineered version of the SUMO fusion platform ideal for mammalian and insect expression due to its resistance to endogenous SUMO proteases found in all eukaryotic systems.
- SUMO Protein Expression Services
Bacteria
SUMOpro and SUMOstar tag systems for E. coli expression
SUMOstar Mammalian
Mammalian host expression systems
Advanced Therapeutic
Yeast
SUMOstar and SUMOpro3 for Pichia pastoris and Saccharomyces cerevisiae
Insect
SUMOstar for insect cell expression
- Bacterial Expression
The SUMOpro tag system is based off our SUMO protein expression platform to maximize the yield of soluble and functional proteins with a native N-terminus. Precise tag removal relies on LifeSensors’ highly specific SUMO proteases, which recognize the tertiary structure of the tag at the N-terminus of the partner protein and cleaves the junction at discrete residues outside the protein sequence.
Therefore, SUMO proteases are superior to other proteases commonly used in prokaryotic expression systems that may non-specifically cleave within the protein of interest or leave extra residues behind. By linking a His6 tag to both the SUMOpro fusion tag and SUMO protease, the system allows efficient and rapid purification of untagged proteins using immobilized metal affinity chromatography. SUMOpro fusion, thus, makes purifying your protein easy and affordable.
LifeSensors offers both yeast (SUMOpro) and human (SUMOpro3) tags with corresponding SUMO proteases 1 and 2, respectively for Bacterial Expression.
Service Highlights
- Convenient, directional cloning of your gene-of-interest in frame with the SUMOpro and SUMOpro3 tag(s)
- Dramatically enhances recombinant protein expression, reducing costs
- Increases protein solubility
- Precise tag removal following protease cleavage
- Yeast Expression
Yeast, especially Pichia pastoris and Saccharomyces cerevisiae, are also commonly used for protein expression. LifeSensors offers both SUMOstar and SUMOpro3 systems for secretory expression in Pichia, while SUMOstar is available and recommended for Saccharomyces intracellular expression. SUMO Protease 2 is compatible with SUMOpro3 expression while SUMOstar Protease is designed to work with SUMOstar
Service Highlights
- Eukaryotic expression for proper folding and wider domains
- Efficient secretion into media for Pichia
- Increases protein solubility, Fast growth, high yields
- Precise tag removal following protease cleavage
- SUMOstar Insect Expression
Insects like many eukaryotic organisms possesses endogenous SUMO proteases that will cleave a protein expressed with a traditional SUMO tag. The insect cell protein expression system, typically using the Baculovirus Expression Vector System (BEVS), involves cloning a target gene into a plasmid vector that generates a bacmid. This bacmid is used to create and amplify recombinant baculoviruses in insect cells such as SF9 expression a target protein. LifeSensors SUMOstar system is recommended for this application.
Service Highlights
- SUMOstar brings SUMO expression benefits to Insect cells
- Dramatically enhances recombinant protein expression, reducing costs, BEVS supported for high yields
- Lower cost than mammalian, suitable for structural studies
- Especially attractive for therapeutic protein production
- SUMOstar Mammalian Expression
While LifeSensors offers both yeast (SUMOpro) and human (SUMOpro3) tags with corresponding SUMO proteases 1 and 2, respectively, in mammalian expression endogenous SUMO proteases can cleave the protein, preventing expression and purification.
SUMOstar is designed to prevent the above issue by using an engineered SUMO tag to prevent cleavage and preserve the enhanced protein production properties of SUMO. Isolated SUMOstar fusions from eukaryotic cells can be cleaved efficiently via SUMOstar protease in vitro.
Service Highlights
- SUMOstar prevents cleavage in eukaryotic cells preserving key properties for enhanced expression
- Dramatically enhances protein expression and solubility, reducing costs in costly mammalian cells
- SUMOstar protease enables removal of tag to generate native N-terminini
- SUMOstar Advanced Applications
Service Highlights
- Large-scale production of biotherapeutics
- Precise, inducible, tag removal to preserve N-terminal residues for biological activity (e.g. cytokines)
- Enhanced secretion of drug targets such as recombinant antibodies, insulin, growth hormones, and stimulating factors
Application of SUMO System across the Biological Spectrum
SUMOpro technology is ideal for enhanced expression, increased solubility, and native folding in E. coli. The SUMOstar system presents similar advantages for native protein expression in mammalian cells.
Chemical synthesis of polypeptides is prone to errors. Not all chemically synthesized peptides fold correctly, and yield of polypeptides decreases along with the length of the polypeptide.
E. coli provides an attractive platform to express animal vaccines at a low cost. The SUMO system provides the capability to increase yield and serves as a tool to rapidly purify the antigen.
Although mammalian cells have proven to be workhorses for antibody production, engineered single chain antibodies (Nanobodies) are especially difficult to manufacture in mammalian cells. SUMOpro is an inexpensive method for Nanobody production.
Despite the strong promoters, many genes are not expressed well in recombinant adeno associated viruses (rAAV). LifeSensors has established a human SUMOstar based system that allows under expressed or non-expressed genes to be delivered as SUMOstar fusion between rAAV ITR containing plasmids.