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Protein Expression and Purification Services
Baculovirus Protein Expression
服务简介

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.

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

  Bacterial Insect Cells(Baculovirus) Mammalian
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.
Cell type E. Coli Sf9, HighFive Mammalian Cell Gene Expression System (Human HEK293 suspension cells)
Expression level Low-Medium-High Low-High Low-Medium-High
Cell doubling time Rapid (30 mins) Slow (18-24 hr) Slow (24 hr)
Timeline 3 weeks 6 - 8 weeks 3 - 6 weeks
Post Translational Modifications
Protein Folding Not reliable Reliable Reliable
Glycosylation No Simple Complex
Phosphorylation No Yes Yes
Acetylation No Yes Yes
Acylation No Yes Yes
γ-carboxylation No No Yes
服务内容

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: 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.

服务特色

We’ve been fueling successful studies with custom-produced proteins since 2013 and look forward to delivering high-quality proteins to help you achieve your study objectives.

Tanda’s protein production services are:

 

1. Comprehensive—we can help you with everything from gene synthesis to protein purification, and beyond. Our services include desalting, aliquoting, lyophilization, and conjugation.

 

2. Fast—you’ll get your bacterially-expressed protein in 3 weeks, insect-expressed protein in 6-8 weeks, and mammalian-expressed protein in 3-6 weeks.

 

3. Easily scalable—once the protein has been purified for the first time, we can scale up production to your specifications.

 

4. High quality—we typically achieve >95% purity as determined with SDS-PAGE.

 

5. Risk free—we run projects in phases. You will be notified of the progress of each phase. You are only billed for milestones successfully completed.

 

6. Affordable.

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