BioActive™ Receptor Protein Production Services

Seattle Genova has comprehensive experience in generating recombinant proteins for research and drug discovery needs. We give complete services from gene cloning and synthesis to protein characterization. Each project is formulated with checkpoints that can give you decision-informing data before proceeding to the next step. We are dedicated to enabling maximize the success rate of your research project. We have developed very specialized protocols to make sure the produced proteins will maintain their biological activities. And our well-developed bioassays can ensure comprehensive activity validations before we release the proteins.

Receptor proteins are found within the cell surface membrane, nucleus membrane or other cellular organelle membrane. They can bind to corresponding ligands to begin cellular signalling pathways. For cell surface receptors, like receptor tyrosine kinases, interleukin receptors and receptors of growth factors, are subdivided into three domains, an extracellular domain,a transmembrane domain and intracellular domain. Receptor proteins construct the largest family of biological targets.

For instance, GPCR (G protein-coupled receptor) is the target of extra than 50% of current drugs. GPCR family members share a different structure with seven-transmembrane domains. Receptor tyrosine kinases are individual transmembrane proteins. They are key regulators for normal cellular procedures and are also comprised in developing many kinds of cancer. The extracellular part of receptor tyrosine kinases is reliable for binding to growth factors and cytokines. The intracellular domain has kinase action. Many researchers have concentrated on developing therapeutic drugs for both extracellular and intracellular domains of receptor tyrosine kinases.

Large Scale Production

The expression vectors are prepared with endotoxin-free plasmid extract kits and are transfected into 2 litres of E. coli or 500 ml of mammalian or insect suspension cells.

Purification 

After the cell culture period, the target protein is purified via affinity chromatography. The concentration and purity of the purified protein are determined using a bicinchoninic acid (BCA) assay and SDS-PAGE with Coomassie blue staining, respectively. The protein is desalted into 0.2 µm filtered 1x phosphate-buffered saline (PBS), pH 7.4. The protein can then be aliquoted, lyophilized, and labelled upon request

Additional Services

Bulk production (optional)

Large-scale gene expression and purification based on the customer's desired amount of purified protein.

Low Endotoxin

Each new production lot of protein is examined for endotoxin using the Limulus Amoebocyte Lysates (LAL) assay. Our definitive endotoxin specification is an industry-leading <0.1 EU/ug.

Highlights

Accurate -All sequences are verified by sequencing and ensured error-free.

Flexible -Synthesize and clone any DNA sequences into our definitive vector or your vector of choice.

High purity -Generate proteins to meet industry-leading purity specifications.

Long-term storage feasibility -Proteins are provided in a lyophilized form which ensures extra long terms of storage without significant protein mass or activity loss.

Talented Staff -Technical support is given by the very scientists that develop thee assays.

Deliverables

➢SDS-PAGE Data

➢WB data

➢The bioactivity Long-term data 

➢The purified proteins are also shipped to our customers for them to test internally

References

1.Alberts B, Bray D, Hopkin K, Johnson A, Lewis J, Raff M, Roberts K, Walter P (2014). Essential Cell Biology (Fourth ed.). New York, NY, USA: Garland Science. p. 534. 

2.Gotti, Cecilia; Marks, Michael. J.; Millar, Neil S.; Wonnacott, Susan (16 September 2019). "Nicotinic acetylcholine receptors (version 2019.4)". IUPHAR/BPS Guide to Pharmacology CITE. 2019 (4).