1. Do you offer protein expression in other expression systems?
No, only in mammalian cells. They have the advantage of full posttranslational modifications of the expressed protein
Frequently asked questions
top2. Which cell type do you use for expression?
HEK293 and CHO cells.
3. Do you use adherent or suspension cells?
We use HEK293 and CHO cells that are adapted to growth in suspension, but we can use adherent cells for protein expression if necessary.
4. How do you cultivate your cells and how much volume can you handle?
We use shaker cultures and wavebag technology. We currently can handle up to 10 liter culture volume.
5. Do you produce membrane proteins ?
No, we currently focus on secreted proteins (i.e. antibodies or cytokines) or secretable proteins with artificial secretion leader.
6. What does the BestExpresser and Best-in-Class service mean?
BestExpresser means that we synthesize different versions of one gene that has been optimized with different optimization strategies (i.e. wt, standard optimized and gene specific optimization), subclone these genes into the same expression vector and compare the three different versions in an expression test (i.e. western blot) and deliver all three constructs and a report with detailed information on the results (gene synthesis and subcloning not included).
Best-in-Class means that we synthesize different gene variants that lead to different protein variants, subclone these genes into the same expression vector and compare the different versions of your protein in an expression test (i.e. Western blot) and deliver all constructs and a report with detailed information on the results (gene synthesis and subcloning not included).
7. Is the price for the generation of the expression vectors included in the cell lines and protein services?
No, the prices are only for the cell line and protein services. All gene synthesis, subcloning and plasmid preparation work is not included. If desired, the expression vectors that we produce are sent upfront and we keep a part of the DNA for the cell lines and protein service work.
8. Can I send you my own expression construct for the production of protein?
Yes, that's possible if you have tested your construct for expression and send us results of your expression analysis; e.g. a coomassie picture with a rough protein quantification and information how much cell culture you used for production. However, we recommend that GENEART produces an expression optimized gene subcloned into a vector of your choice and does the production of your protein from that optimized construct.
9. I want Xmg of my protein. What is the price?
Before we can tell you a guaranteed price, we need information on the productive performance of your protein. This information will be gained in a Genes to Proteins "pilot" production where we define the expression and purification performance. After the Genes to Proteins "pilot" service, we can provide you with a binding quote for a specific protein amount.
10. What price range do you have, can I have an overview on the prices for the different services?
No, since cell line production and protein production projects are so different and individual, each project is priced separately. Please define a specific project in an email to: protein@geneart.com. We will come back to you with a price quote.
11. Can I have an overview on your services?
Yes, we'll provide you with a service overview if you send an mail to: protein@geneart.com. Please specify what service you are most interested in (Genes to Expression, Genes to Proteins, Genes to Cell lines).
12. What purification methods do you use?
We use all available methods for purification, e.g. affinity tag for purification, ProteinG/A (for antibodies or their derivatives) or combinations of IEX/HIC and GF chromatography.
13. If you use an affinity tag, can you cleave it off?
Yes, we use tags that can be cleaved off after purification and removed from the purified protein (e.g. by introducing a thrombin or TEV cleavage sequence during gene synthesis, some aa may be left at the protein after cleavage).
14. What is ment by "uncloned stable cell line" or "stable cell pool"?
After transfection, the cells are cultivated until only stably transfected cells are growing in the culture. For the "polyclonal" service, we check for transgene expression and deliver this cell pool of uncloned, stable transfected cells.
For the "clonal" service, we subclone several 100 different cell clones from a stable cell pool, check for transgene expression and deliver clonal cell line(s) expressing the protein of interest.
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