Below are some frequently asked questions about our Protein Production services. Should you have any questions other than listed below, feel free to contact us at firstname.lastname@example.org and we will be happy to clear your thoughts.
Currently we offer E.coli, mammalian, Yeast, and Baculovirus (insect) expression systems. We offer extensive production and purification services, including tag-removal, lyophilization, endotoxin removal, fermentation, N-terminal Sequencing and QC testing. Our gene synthesis services also allow us to synthesize and subclone your gene to get it ready for protein production.
E.coli expression is by far the most cost-efficient and consistent production method. We have the capabilities to scale up from milligrams to grams. Although E.coli do not incorporate post-translational modifications into the recombinant protein, many E.coli produced proteins do show activity of the native protein (our robust catalogue of active recombinant proteins is evidence of this). However bacterial cells produce endotoxins, so if you plan to use the protein in any animal cells or assays, endotoxin removal is recommended.
Baculovirus (insect) expression is ideal for producing mature, folded, and processed recombinant proteins. In contrast to E. coli, insect cells are able to incorporate post-translational modifications, they are also particularly well-suited for expressing secreted proteins.
Proteins produced in mammalian cells have the advantage of containing full post-translational modifications (e.g. proper folding, glycosylation, phosphorylation, etc) of the expressed protein. However, mammalian expression levels tend to be lower, so the cost is higher and achieving large amounts of protein (>10mg) may be difficult.
We have codon optimization and gene synthesis services, so if you just have your sequence as a FASTA or NCBI entry, we can use that to get started. Or if you already have your cDNA in an expression vector we can use your provided construct as well. We would also request that you complete our protein order form so that we can understand your requirements.
Yes, we will use an affinity tag (e.g. HIS or GST) to purify your protein. For difficult to express proteins, we may even try alternate tags (including SUMO or thioredoxin) to aid in solubility and expression. If needed, we can definitely cleave the tag and provide tagless protein as the final product.
Our experienced protein team will do what it takes to purify almost any protein. However, for best success we recommend keeping the target protein <100kDa and avoiding proteins with too many transmembrane domains. We also recommend removing any signal peptides that may be attached to the N-terminus of the protein, as these can inhibit protein expression in recombinant systems.
For all projects we include free SDS-PAGE analysis and protein concentration determination. If required we can also provide HPLC, MS, and N-terminal sequencing for modest additional fees.
Yes, for E.coli expression systems we are capable of producing large amounts of protein in a single lot (up to gram amounts!). For these large scale projects, we will typically start with a small-scale pilot expression and purification, where we will purify ~5mg of protein and send it to you for testing. If it meets your needs we can then scale up the production.
In most cases our production facilities meet or exceed GMP standards. In addition, we are ISO9001:2008 certified to supplement our quality commitment.
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