Protein Production Services

Start your experiments off right with pure, stable, and active proteins reliably expressed and purified by RayBiotech.

Our comprehensive protein production services cover every step of the workflow, from gene cloning and sequence validation to expression and purification. We can also conjugate your protein to a moiety of interest—such as a fluorophore, biotin, or streptavidin—and perform downstream processing activities such as protein desalting, lyophilization, vialing, and labeling.

protein-molecule

Why You Should Choose Our Protein Production Services

We’ve been fueling successful studies with custom-produced proteins since 2006 and look forward to delivering high-quality proteins to help you achieve your study objectives.

RayBiotech’s protein production services are:

  • Comprehensive—we can help you with everything from gene synthesis to protein purification, and beyond. Our services include desalting, aliquoting, lyophilization, and conjugation.
  • Fast—you’ll get your bacterially-expressed protein in 3 weeks, insect-expressed protein in 6-8 weeks, and mammalian-expressed protein in 8 weeks.
  • Easily scalable—once the protein has been purified for the first time, we can scale up production to your specifications.
  • High quality—we typically achieve >95% purity as determined with SDS-PAGE.
  • Affordable.

See What Our Protein Production Service Can Do For Your Research

We Offer a Range of Protein Expression Systems to Support Different Research Needs

Bacterial Baculovirus Mammalian
Cell type E. Coli Sf9, HighFive TruExp Mammalian Cell Gene Expression System (Human HEK293 suspension cells)
Expression level Low-Medium-High Low or High Low-Medium-High
Cell doubling time Rapid (30 mins) Slow (18-24 hr) Slow (24 hr)
Cost $ $ - $$$ $
Timeline 3 weeks 6 - 8 weeks 8 weeks
Post translational modifications (PTMs) None Yes, but different types and patterns from mammalian systems with no sialic acid or gamma-carboxylation Yes
Summary Bacterial gene expression is a fast and cost-effective approach for producing recombinant proteins. Baculovirus insect cell gene expression is easy to scale up from the laboratory to a large-scale production system. However, while the expressed proteins have abundant PTMs, the types and patterns of modifications are different from those seen when the same protein is expressed in a mammalian system. Mammalian gene expression is the best option for producing native conformations and PTMs of mammalian proteins.

See the Power Of Our Proprietary TruExp™ Mammalian Cell Gene Expression System

TruExp is RayBiotech’s state-of-the-art mammalian cell-based expression system developed for producing recombinant proteins with native conformations and PTMs. TruExp is an excellent option for proteins that have a high molecular weight or are difficult to express.

We recommend TruExp Mammalian Cell Gene Expression for:

  • Complex, abundant PTMs—see Figure 1
  • High expression—typical yield ranges from 2 - 355 mg purified protein per liter of culture (Figure 1, Table A).
  • Secretion—expressed protein is secreted into the culture medium, facilitating high purity levels (in contrast to recovering protein from highly contaminated lysate fractions) (Figures 1 & 2).
  • No biohazard—serum-free medium and animal source-free materials eliminate biohazard risk.
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Figure 1. The TruExp Mammalian Cell Gene Expression System delivers high protein expression levels. Six human genes (H1-6) and 3 mouse genes (M1-3) were expressed by the TruExp expression system. Cell culture supernatants (10 µL) were applied to 12% SDS-PAGE (A) and Western blot probed with anti-his tag monoclonal antibody (B). Except for genes H6 and M2, all overexpressed genes display additional secreted protein bands (A, red arrows). All genes were expressed, with some displaying abundant post-translational modifications, evident by increased molecular weights compared to the predicted sizes (B, yellow arrows). M = protein standard markers (kDa).

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Figure 2. Proteins expressed in the TruExp Mammalian Cell Gene Expression System are full-length and highly pure. Affinity-purified recombinant proteins from culture supernatants of transfected mammalian cells exhibit high purity and intact protein bands, even at MW larger than 100 kDa. M = protein standard markers (kDa).

Ready to see what our protein production service can do for your research?

How Our Protein Production Service Works

Phase 1: Gene synthesis (usually 2-4 weeks)
The target gene is synthesized, using a sequence that is codon optimized for the expression system.

Phase 2: Gene cloning (1 week)
The target gene is cloned into RayBiotech OVER-expression vectors with an N-terminal signal peptide for protein secretion and an N-terminal or C-terminal 6x Histidine (His) tag for protein affinity purification. The recombinant construct is confirmed by DNA sequencing.

Phase 3: Small-scale gene expression (1-2 weeks)
The plasmid cDNA is extracted from the cells used for cloning and transfected into 5 mL of the expression cells. We examine the target protein in the cell lysate and supernatant via SDS-PAGE and/or western blotting using an anti-His tag antibody to evaluate gene expression levels, protein expression levels, protein secretion, and protein solubility. If the protein does not meet your expectation, the service can be stopped after this phase.

Deliverable: Electronic SDS-PAGE and/or western blotting results.

Phase 4: Large-scale gene expression and purification (2-4 weeks)
The plasmid cDNA is extracted from the cells used for cloning and transfected into 2 liters of the expression cells (3 million cells per ml). The cells are cultured and the target protein is purified via its His tag using immobilized metal ion affinity chromatography (IMAC). 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 (additional lead time may apply).

Deliverable: All the protein purified in this phase.

Phase 5: Bulk production (optional)
Large-scale gene expression and purification based on your desired amount of purified protein.

Deliverable: All the protein purified in this phase.

Frequently Asked Questions

Still have questions?

We guarantee that your target gene-of-interest is correctly synthesized using DNA sequencing.

We cannot guarantee that your protein will be expressed since expression level cannot be predicted. This is why a small-scale gene expression test to determine protein expression is performed in Phase 3. However, membrane proteins are often poorly expressed and purified. Therefore, we recommend that only soluble domains are expressed.

You only need to pay for the work that is performed. That is, if the results in Phase 3 show that the expression failed and you choose not to continue to Phase 4, you do not need to pay for Phase 4.

The signal peptide causes the protein target to be secreted out of the cell and into the serum-free culture media where the protein content is minimal. This results in a purer purified protein since there are fewer proteins in the culture supernatant (compared to the cell lysate) to nonspecifically bind to the affinity purification column. For example, protein purity > 95% is routinely achieved with secreted proteins, whereas protein purity > 50% is often obtained with non-secreted proteins.

Sometimes the protein target will not be secreted even with the signal peptide. It is likely that, in the protein tertiary structure, the signal peptide is occluded.

The signal peptide is 25 amino acids long. It is often cleaved by proteases in cell culture, but we cannot guarantee that the signal peptide will be cleaved. Upon request, we can exclude the signal peptide in the expressed protein.

Yes. If you have already expressed your target protein successfully using your expression vector, please provide the expression conditions that you would like us to use. We can then go to Phase 4 directly. If you have not confirmed that the gene is expressed in your expression vector, we will perform a small-scale expression test in Phase 3.

It depends. Please provide us with the gene sequence and, at a minimum, 100 nucleotides flanking the gene at the 5' and 3' ends. This information is necessary because we will use restriction enzymes to cut the gene out of your plasmid and ligate it into ours (i.e., cloning). If we can clone your gene into our vector, then Phase 1 is not necessary. If the gene and flanking sequences are incompatible with any restriction digest approach, then we cannot use your gene and will have to start at Phase 1.

Also, it is important to consider whether your gene is codon optimized for the expression system.

Yes. Please indicate this in the quote request form.

We can add a cleavage site between the tag and target protein.

For example, we can add a tobacco etch virus (TEV) site between the His tag and protein-of-interest. After the His-tagged protein is purified, a His-tagged protease could be added, cleaving the His tag from the target protein. The tagless protein could then be purified from the His tag and His-tagged TEV protease using IMAC.

If the protein was expressed and purified previously, you do not need to pay for Phases 1 - 4 again. You would simply complete the quote request form for the desired amount of purified protein (Phase 5). Please make sure to include the previous sales order or invoice number in the Project Description section.

Protein Name Yield (mg/L culture
Serpin B4, Serpin Peptidase Inhibitor SCCA2 335.7
Calcitonin CT 160.8
14-3-3 Protein Epsilon YWHAE 103.1
N-terminal of the prohormone BNP NT-proBNP 99.5
Lipocalin-2 NGAL 69.1
a-Fetoprotein AFP 63.2
Brain natriuretic peptide 32 BNP-32 51.2
Ubiquilin-1 UBQN 41.3
Carcinoembryonic Antigen CEA 39.8
Receptor Tyrosine-Protein Kinase erb-3 erbB-3 37.5
Epidermal Growth Factor Receptor EGFR 23.5
Pepsinogen II, Pepsinogen C PGC 18.6
Lymphocyte Activation Gene 3 Protein LAG-3 16.9
Receptor Tyrosine-Protein Kinase erbB-2 Erb/Her2 16.7
Hepatocyte Growth Factor Receptor HGF R 14.6
Annexin A2 ANXA2 11.8
Golgi Membrane Protein 1 GP73 5.5

Still have questions?

Featured Resources

Get additional information to help you advance your studies from articles on the science underlying antibody-based techniques to video protocols, application notes, and more.

    Proteins & Peptides
    Recombinant Proteins – Comprehensive Gene To Protein Service
    Proteins & Peptides
    Recombinant Proteins in Research
    White Paper
    Pushing the Limits of Multianalyte Immunodetection

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