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BioActive™ Biological Target Protein Production

BioActive™ Biological Target Protein Production

Seattle Genova offers our customers excellent protein production services that keep the bioactivities of your proteins just as 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.

Biological target is very significant to drug discovery, drug development and disease diagnosis. Based on published measures, there are nearly 1100 host biological targets and 200 pathogen biological targets. Among them, about one-third are therapeutic affect-mediating targets (also recognized as drug targets). 

According to their purposes in signalling pathways, host biological targets can be categorized into receptors, enzymes, transporters, and other molecules. The best-known receptor family is G protein-coupled receptors (GPCRs), pursued by receptor tyrosine kinases and ligand-gated ion channels; while the most popular enzyme family is hydrolases, then oxidoreductases and transferases. 

Biological target can be utilized in the pharmaceutical industry and academic biomedical research for immunomodulation, inflammation, cardiovascular diseases, central nervous system (CNS) and cancer etc.

Cancer-type target protein

Targeted Protein for liver cancer

Targeted Protein for kidney cancer

Targeted Protein for lungs cancer

Targeted Protein for breast cancer

Targeted Protein ovarian cancer

Targeted Protein for pancreatic cancer

Targeted Protein for colorectal cancer

Targeted Protein for leukaemia 

Targeted Protein for prostate cancer

The word "biological target" is often utilized in pharmaceutical research to define the native protein in the body whose activity is modified by a drug arising in a specific effect, which may be a desirable therapeutic effect or an unwanted effect. In this context, the biological target is often related to a drug target.

 The most familiar drug targets of nowadays marketed drugs include:

Proteins

 ◇G protein-coupled receptors (target of 50% of drugs)

 ◇enzymes (especially protein kinases, proteases, esterases, and phosphatases)

Ion channels

 ◇ligand-gated ion channels

 ◇voltage-gated ion channels

Production Process

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.

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.

Transfection: 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. 

Protein Analysis:The target protein in the cell lysate and supernatant is analyzed via SDS-PAGE and/or western blotting using an anti-tag antibody.

Protein Validation: 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.


Large-scale Expression & Purification Process

The expression vectors are prepared with endotoxin endotoxin-free tract kits and are transfected into 2 litres of Elites 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.


Features 

Advanced equipment and technique

Experienced scientific team

High Purity

Our proteins meet industry-leading purity specifications.

Biological Activity

Proprietary methods for accurate protein for cancer ensure biologically relevant proteins.

Lot-to-lot Consistency

Multiple lots are created for proteins, all with matching specifications.

Lower Endotoxin

Our standard endotoxin specification is an industry-leading <0.1 EU/ug.

Long-term storage feasible

Proteins are delivered in a lyophilized form which ensures extra-longg terms of         storage.

Standard Quality control

Highly reliable and reproducible result

Quality one-stop service


Deliverable

SDS-PAGE data

WB data

Bioactivity Validation data

Purified protein


References

1.Raffa RB, Porreca F (1989). "Thermodynamic analysis of the drug-receptor interaction". Life Sciences. 44 (4): 245–58.  

2.Moy VT, Florin EL, Gaub HE (October 1994). "Intermolecular forces and energies between ligands and receptors". Science. 266 (5183): 257–9.  

3.Srinivasan, Bharath (March 2022). "A guide to enzyme kinetics in early drug discovery". The FEBS Journal.  







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