Recombinant Protein Production
Recombinant protein production is a prerequisite and essential component of both small molecule or large molecule drug discovery programs. At BioMetas, we have a wealth of expertise in recombinant protein generation, offer custom recombinant protein production services from a range of expression systems (E.coli, insect cell, mammalian) along with other protein services like membrane protein services, protein labeling, protein refolding, etc.
Our services include all the steps from gene synthesis and expression vector construction to pilot protein production, large-scale protein expression with fast project processing according to the highest quality standards. We devote ourselves to provide high-quality reagents to accelerate your discovery.
Services
- Bacterial Expression
- Insect Cell Expression
- Mammalian expression
- Membrane protein production
- Protein labeling
- Protein refolding
Advantages
- Versatile Expression System
- Difficult-to-expression protein generation experts
- Quality and quantity guaranteed
- Competitive pricing
- Fast turnaround: 2-4 weeks from gene to pilot production
Protein Purification
- Affinity chromatography
- Ion exchange
- GST/HIS-tagged purification
- Hydrophobic interaction chromatography (HIC)
- Size-exclusion chromatography (SEC)
Quality Control
- SDS-PAGE
- Western Blot
- Endotoxin determination
- ELISA
- Activity assays
Bacterial expression services
Rapid and cost-effective production of large amounts of proteins usually requires bacterial expression system. The technology development of this system is relatively perfect, which is suitable for the expression of proteins from different species, and has great advantages for the production of proteins with small molecular weight. But there are some problems with the system, such as: The poor compatibility between some expression vectors and strains may result in no or low expression of target proteins; The expressed target protein is insoluble and easy to form inclusion bodies;
Proteins produced from bacterial expression systems usually contain endotoxins, which are difficult to be effectively removed by conventional methods.
In order to ensure the production and quality of each recombinant protein, BioMetas has developed a complete toolbox to tackle any such challenges and to provide best-in-class protein engineering services, providing one-stop service from sequence design, plasmid construction, cell expression, to protein purification, as well as endotoxin removal and other additional services.
Our scientists have rich experience in prokaryotic expression and purification. By selecting different host expression systems (Escherichia coli and Bacillus subtilis), different expression vectors containing multiple fusion labels (His, Trx, GST, Sumo, Flag, MBP, etc.), different cell lines (BL21,Rosetta,C41,WB800, etc.), optimizing the expression conditions, and refolding, etc., greatly solve the problem of prokaryotic expression system.
- Highlights
- Multiple vectors and hosts to ensure soluble protein expression and protein activity
- Purity>95% for most cases
- Endotoxin level can be controlled as low as 1EU/mg
- Turnaround time : 3-5 weeks
Case Study
2-subunit complex
Quinary complex (E.coli)
The traditional approach to target based small molecule drug discovery strongly depends on the in vitro screening of disease relevant proteins as single polypeptide chains. But in vivo , most of the disease targets being pursued by the pharmaceutical industry are not proteins that act in isolation. They are rather acting as part of multi-protein complexes. Determination of the subunit composition, stoichiometry and status of posttranslational modifications ideally are the starting point for cloning, expression and purification of the disease target as a relevant protein complex in the desired functional state. BioMetas provides many choices of expression systems for multiprotein complexes generation.
Above are two example of protein complex produced in e.coli system.
Insect cell expression services
Biometas provides the high-quality recombinant protein expression services using our in-house baculovirus–insect cell system from gene synthesis to protein production. Compared with bacterial expression system, this platform supports improving protein solubility of intracellular and secreted proteins with complex post-translational modifications. In addition, its high protein expression levels compared to other higher eukaryotes system renders it an excellent choice for difficult-to-express proteins, including but not limited to cytotoxic proteins, larger proteins, protein complexes and membrane proteins. We highly recommend this cost-effective eukaryotic protein expression system for eukaryotic protein production. Combining our long-standing expertise in protein production and a complete platform, Biometas optimize all the steps and boost the yield of your insect cell expression.
- Highlights
- Long-standing expertise
- Wide variety of cell lines and expression vectors
- Codon optimization for insect cell expression
- Several optimization parameters: insect cell line, Virus titration, multiplicity of infection and incubation time
Case Study
Protein kinases remain one of the most interesting targets for small molecule targeted drug discovery. High quality protein is fundamental to set up in vitro assays. Though most of kinases are commercially available, sometimes it’s cost-prohibitive especially when large amount protein is required for a specific study. Below is case study of producing a well-known kinase protein by baculovirus system. 10mg of target protein was produced in 6 weeks with desired purity and activity.
Mammalian cell expression services
Mammalian cell is an important expression system in the gene engineering pharmaceutical research. It has highly sophisticated mechanism of protein folding and post-translational modification, the recombinantly expressed protein has correct folding and modification, thus the conformation is much closer to human. Because the product is nearest to the natural protein molecule of higher organisms in molecular structure, physical and chemical properties and biological function, mammalian cell has become the preferred host for producing therapeutic recombinant proteins.
BioMetas provide protein production by transient expression in both HEK293 and CHO cell lines. We are able to carry out high throughput and high yield production for recombinant antibodies and proteins. The transiently expressed products can be applied to antigen generation, antibody discovery, protein structure and function research, etc.
- Highlights
- From transient to stable protein expression
- Proprietary mammalian high performance system
- Diverse purification methods, e.g. affinity chromatography, SEC, IEX, HIC, etc.
- Short turnaround time and competitive price
Case Study
Addition of an affinity tag is a useful method for purifying a target protein from the total cell lysate. In general fusion- protein tags can be classified into two main groups: solubility tags (GST, MBP, NusA, Trx and SUMO) and affinity tags (Poly His, Flag, HA, Strep II). And tags can be place on protein’s C-terminus or N-terminus. Below is a case study where we tested two tags (MBP and flag) on 12 proteins to investigate their impact on protein’s yield and quality.