Transient Expression of functional proteins in suspension cultured CHO or Hela cells
- Strong eCMV promoter to produce and purify up to 100 mg protein in 4~6 weeks
- Laboratory scale from 100 ml to 3 L of cultured cells.
Pilot Expression
A pilot project will be carried out before starting a large-scale protein production service. Optimal parameters for protein expression will be determined for each individual protein.
- Infection of HeLa cells using previously generated recombinant adenovirus with three different MOI (multiplicity of infection).
- Harvesting infected cells at two time points, typically at 48 and 72 hours post-infection.
Time Required: 2-3 weeks.
Large-Scale Protein Production
Protein expression in CHO or HeLa cells, according to characteristics of the recombinant adenovirus or customer's specifications.
| Service No. |
Production Scale |
Infection of |
| A2P500 |
500 mL |
5x108 cells |
| A2P1000 |
1 L |
1x109 cells |
| A2P2000 |
2 L |
2x109 cells |
| A2P3000 |
3 L |
3x109 cells |
Deliverables
- 1 to 100 mg purified (> 95% purity) His-Fc fusion protein, cleavable by TEV protease or enterokinase.
- QC Data
Required Materials
- Plasmid containing gene of interest plus complete vector sequence in electronic format
- or protein sequence for gene synthesis (with additional price for gene synthesis)
- or accession number
Quotation and Ordering
To request a quotation, please request a quotation form from customer services crm@excellgen.com
To order, please send the completed quotation form to us by email or fax with a formal PO (Purchase Order) or credit card information. You can also submit PO/credit card information by phone.
If submitting samples, please mail them together with a hard copy of the completed quotation form to: Protein Services, Excellgen, Inc. 15601 Crabbs Branch Way, Rockville, MD 20855-2743.
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Excellgen's scientists have average of 15 years experience in molecular biology, protein expression and purification. Our team is proficient in molecular cloning, producing and purifying of recombinant proteins in a variety of expression systems including cell-free protein synthesis, E. Coli, yeasts, baculovirus/insect cells and mammalian cells (CHO and 293). We have ÄKTAExplore and ÄKTAPurifier FPLC systems, ~ 30 large GE Pharmacia prepacked columns (gel filtration, hydrophobic, ion exchange, desalting) and numerous affinity columns. By combining state-of-the-art fementation and ÄKTA FPLC chromatography systems, we can produce quantities of pure recombinant proteins ranging from miligrams to a few grams.
Cell-free protein synthesis, aka in vitro translation, is a powerful approach to synthesize desired protein rapidly. For example, proteins which are toxic to living cells and thus usually can not be easily expressed in cell-based systems, could be easily produced ranging from 100 microgram to miligrams using a cell-free protein synthesis system.
Excellgen offers cell-free protein synthesis services using commercial available cell-free extracts from both procaryotic or eukaryotic cells. The choice of the cell-free protein synthesis system should be determined by: 1), the origin and biochemical nature of the protein to be synthesized; 2), the specifics of the downstream applications.
Advantages
- High success rate of target protein expression
- Natural protein folding with high percentage of active proteins
- Production process is automatable therefore high throughput and scalable
- Suitable for production of proteins that are potentially toxic to cells
- Incorporation of labeled and/or modified amino acids for special assays
Applications
- Protein interaction studies
- Pull-down protein complex
- Protein immobilization
- Protein labeling (biotinylation and isotope labeling)
- Active protein array for drug target validation and lead screening
- Real-time protein array to solve problems in protein chip
- Antigen used for raising polyclonal or monoclonal antibodies
- High density protein chip for antibody screening and validation
Services
- PCR and Subcloning
- Cell-free Synthesis
- FPLC Purification and SDS-PAGE Analysis.
Timeline
- Cloning and subcloning, sequencing: 1-2 weeks (cell-free protein synthesis using PCR template does not need cloning)
- Cell-free synthesis, FPLC purification and QC: 1 week.
Depends on the availability of cDNA to be used for protein synthesis, and the downstream application, we may be able to synthesize protein for your lab within one week starting from cDNA clone.
Reagents used for in vitro protein synthesis
- NEB PURExpress
- Qiagen EasyXpress Protein Synthesis Kit
- Roche ProteoMaster E. coli HY Kit
- Roche RTS 500 Wheat Germ CECF Kit
- Promega TNT Protein Expression
- Invitrogen Expressway™ Cell-Free Expression System
- CellFree Sciences ENDEXT Wheat Germ Expression Kits
- Thermo Scientific Pierce Human In Vitro Expression System
- Stable isotopes from Cambridge Isotope Laboratories
Deliverables
- Purified Protein
- QC Data
- Chromatograms
Quotation and Ordering
To request a quotation, please request a quotation form from customer services crm@excellgen.com
To order, please send the completed quotation form to us by email or fax with a formal PO (Purchase Order) or credit card information. You can also submit PO/credit card information by phone.
If submitting samples, please mail them together with a hard copy of the completed quotation form to: Protein Services, Excellgen, 847 Quince Orchard Blvd, Suite H, Gaithersburg, MD, 20878, USA.
