E coli Protein Service
The advantages of E. coli expression systems include easy operation, low cost, high expression, as well as in-depth research on its genetic background and the diversity of vector strain selection, the use of E. coli to overexpress target genes is still the preferred method at present. However, the codon preferences of different species and the cytotoxicity and solubility of expressed products affect the expression of heterologous genes. With our rich experience, We offer multiple fusion/affinity tags to increase the protein solubility, expression levels, and ease of purification.
| Highlights | Details |
|---|---|
| High Success Rate | Success rate of the soluble protein expression provided using E. coli can be up to 95%. |
| Optional Expression Vectors | NewEast provides a variety of screened and reformed E. coli expression vectors to meet the specific requirements of intracellular and periplasmic; Space secretory expression including the pET series, the pBAD series, the pGEX series; The pCold series. |
| Optional Protein Tags | We use solubilized tags such as SUMO, Trx, GST, and NusA to improve; The hydrophilicity of protein in inclusion body expression. |
| Optional Expression Hosts | NewEast can reform and use more than 10 E. coli expression hosts; Including the standard expression strain, the protein expression enhanced strain, the toxic Protein expression strain, the ultra-low temperature strain for enhancing soluble protein expression, and the other expression strains, to meet the customers’ different requirements. |
| Large-scale Production | 1-4,000 L per batch. |
| One-stop Service | From design and optimize gene, optimize protein expression conditions, purify protein and post processing, to managing of data and report. |
| Step | Description | Timeline |
|---|---|---|
| Plasmid Construction | Gene synthesis with codon optimization & subclone | 2 Weeks |
| Protein Expression Test | Plasmid transformation Exploration of expression condition, expression analysis and identification (SDS-PAGE and/or WB) 200 mL amplification in optimum condition and purification test | 3 Weeks |
| Large-Scale Fermentation & Purification | Scale up protein expression using optimized conditions Protein affinity purification (desired protein amounts and purity) Denaturation and renaturation of inclusion bodies (if required) Tag-free protein (if required), endotoxin removing (if required) Quality control | 3-5 Weeks |
Mammalian Protein Expression Service
Advantage
| Highlights | Summary |
|---|---|
| High activity and high success rate | Proteins expressed in mammalian cells can achieve complex post-translational modifications. |
| Multiple expression system options | Transient transfection expression Stable transfection expression Recombinant antibody expression,and preparation |
| One-stop Service | Design and optimize gene Optimize protein expression conditions Purify protein and post processing Manage data and report. |
Stable Cell Line Construction
| Step | Description | Timeline |
|---|---|---|
| Vector Design | Conducts bioinformatic analysis on target(s) and recommends an expression strategy to each client. | 1 Week |
| Plasmid Construction | Gene optimization and synthesis, Vector construction | 2-3 Weeks |
| Identification of polyclonal stable cell lines | Vector transformation, screening and identification identification of positive cell clones | 2-3 Weeks |
| Vector transformation, screening and identification identification of positive cell clones | Cell subclonal screening to screen out monoclonal cell lines with high expression levels and stable performance | 3-4 Weeks |
| Stable cell line quality test | Perform performance verification according to customer requirements, such as protein activity, expression level, cell passage stability, etc. | 1-2 Weeks |
Yeast Protein Expression Services
Yeast expression system is one of the most economical eukaryotic expression system for scale up production of both secreted and intracellular proteins that require post-translational modifications
Yeast protein expression system is the most economical and efficient expression system among eukaryotic protein expression systems, which combines the advantages of prokaryotic expression system and eukaryotic expression system. Because of its fast growth rate, low culture cost and clear genetic background, it has been widely used in industrial microorganisms. The yeast expression system can correctly process, modify and fold post-translational proteins, so it is often used for the expression of heterologous functional proteins in eukaryotes. Among them, the Pichia pastoris expression system is more widely used. Compared with other systems, it has unique advantages, such as strong promoters FLD1, GAP, AOX1, etc., which can make the expression of heterologous proteins lasting and efficient.
Advantage
| Highlights | Summary |
|---|---|
| High activity and high success rate | According to the codon preference, to optimize the target gene, Effectively improve protein solubility and expression |
| Multi-carrier selection scheme | According to the characteristics of different proteins Vectors suitable for extracellular secretion expression can be provided, vectors that can also be expressed intracellularly |
| Optimization of induction conditions | By optimization of Conditions such as methanol concentration and induction time , we can effectively improve the expression of protein. |
| Extra Services | Tag removal Endotoxin removal |
Stable Cell Line Construction Service
| Process Step | Description | Timeline |
|---|---|---|
| Vector Design | Conducts bioinformatic analysis on target(s) and recommends an expression strategy to each client. | 1 Week |
| Plasmid Construction | Gene optimization and synthesis, Vector construction | 2-3 Weeks |
| Identification of polyclonal stable cell lines | Vector transformation, screening and identification identification of positive cell clones | 2-3 Weeks |
| Screening of monoclonal stable cell lines | Cell subclonal screening to screen out monoclonal cell lines with high expression levels and stable performance | 3-4 Weeks |
| Stable cell line quality test | Perform performance verification according to customer requirements, such as protein activity, expression level, cell passage stability, etc. | 1-2 Weeks |