BioActive™ Protein Production Services

Seattle Genova’s BioActive^™platform is aiming to offer our customers with excellent protein production services that keep the bioactivities of your proteins just as what they were in nature. To achieve this, we can offer both nature protein purification services and recombinant protein production services. 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.

Working with a variety of protein expression systems, we are able to rapidly generate bespoke protein reagents to meet customer needs. Backed by years of expertise, we have successfully produced of a variety of proteins for many biological studies.

Phase 1: Gene synthesis

We do sequence analysis and codon optimization for the expression system that will be used. The designed sequences are then synthesized to generate a DNA template.

Phase 2: Gene cloning

The target gene is cloned into expression vectors for the mammalian, insect or E.coli cells. Multiple N-terminal signal peptides for protein secretion and N-terminal or C-terminal tags for protein affinity purification are available to choose from. The recombinant construct is confirmed by DNA sequencing.

Phase 3: Pilot-scale gene expression

The plasmid cDNA is extracted from the cells and transfected into 100 ml of E. coli or 50 ml of mammalian or insect suspension cells. The target protein in the cell lysate and supernatant is analyzed via SDS-PAGE and/or western blotting using an anti-tag antibody.

Phase 4: Bioactivity Testing

We can develop bioassays to validate the activity of the proteins we prepared. Only those proved to be active are qualified for release. Otherwise, we will start over to try different protocols until the bioactive protein is prepared.

Deliverable: SDS-PAGE, WB and the bioactivity validation data are sent to our customers. The purified proteins are also shipped to our customers for them to test internally.

Phase 5: Large-scale gene expression and purification

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

Deliverable: The customer is sent all the protein purified in this phase.

Phase 6: Bulk production (optional)

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

Deliverable: The customer is sent all the protein purified in this phase.

What can you expect?

High Purity

With complete control over all quality testing, Seattle Genova can generate proteins to meet industry-leading purity specifications.

Biological Activity

Proprietary methods for accurate protein folding ensure biologically relevant proteins and activity can be evaluated per customer’s request.

Lot-to-lot Consistency

Multiple lots are created for Seattle Genova’s proteins, all with matching specifications. Each new lot is tested side-by-side with previous lots to ensure unmatched consistency.

Low Endotoxin

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

Long term storage feasible

Proteins are delivered in lyophilized form which ensures extra long terms of storage without significant protein mass or activity loss.