Mammalian Cell Expression System

Introduction:

The prokaryotic expression system has the advantages of high expression level, simple operation, short cycle, easy large-scale and high-density culture and low cost. For the full-length antibody and glycoprotein biological drug, the folding of expression product polypeptide chain, the disulfide bond, the presence or absence of glycosylation and the type of glycosylation often affect the properties of the synthesis, secretion, biological activity, in vivo stability, and immunogenicity of the expressed product. Compared with other eukaryotic expression systems, the expression of the target gene in mammalian cells is similar to that of the native protein in the type and manner of the glycosylation, and can be correctly assembled into the multi-subunit protein.

Advantages:

• No self-produced endotoxin
• Secretion expression is available
• With a variety of complex N-linked glycosylation, accurate O-linked glycosylation and other post-translational processing
• Close to native protein in the molecular structure, physical and chemical properties and biological functions
• High yield after system optimization: as high as 100 mg/L
• Multi-cell lines, multi-expression methods to improve the protein expression success rate and protein yield
• Price as low as $1280/1mg, delivery time as short as 35 business days

Guarantee:

Risk-free: We do NOT charge if we cannot deliver the protein.

Note: This risk-free custom protein service is only suitable for proteins within 800aa. We will charge according to different steps if you need to express proteins more than 800aa.

Service Process:

Steps	Project	Process		Cusabio Features	Lead Time
1	Plasmid construction	Codon optimization; gene synthesis		Multiple vectors optimization, More options for customers In order to improve the success rate of expression and achieve higher yield, in addition to conventional N-terminal fusion protein expression, we also provide C-terminal fusion protein, which retains the bioactivity of the protein while ensuring high purity.	15-20 business days
		The PCR amplification products are ligated to the vectors e.g. pSec series, pCMV series and pcDNA series vectors
		Transform TOP10 E.coil competent cells
		Obtain the correct recombinant plasmid
2	Small scale expression	Prepare the transfection grade recombinant plasmid in large quantities		Optimization of transfection conditions Set different transfection conditions, select the optimal experimental conditions according to the test results.	9-11 business days
		Transient transfect HEK293, CHO and other cells
		Detect expression products
3	Scale up expression and purification	Scale up the culture cells and transfect		Multi-condition expression scheme According to the protein localization and the best experimental conditions in the small test expression, select different cell lines and different ways of transfection, which can increase the expression quantity, greatly improve the protein expression.	8-9 business days
		Explore different chromatographic conditions including ion exchange, hydrophobic and others by using AKTA, and then determine the optimal purification method.
4	Additional services (Optional)	Charge	Tag removal by restriction digestion	Flexible additional services Customers can flexibly choose from a variety of additional services to their specific needs, e.g. Endotoxin removal, Filter-sterilization, Tag removal, Lyophilization, etc. Some are complimentary, and some require additional charge.	3 business days
		Free	Filter-sterilization; Endotoxin removal; Lyophilization (Note: Lyophilization and Filter-sterilization can not be met simultaneously)		2 business days
5	Quality Control	Testing of purity, concentration, etc. QC report is provided.		Detailed COA report Detailed product data sheet and COA are provided for each project.	3-5 business days
Total lead time					35-45 business days