Mammalian Cell Protein Expression Service
Mammalian expression system are the primary option for the production of recombinant proteins and antibodies in pharmaceutical research. Mammalian cells can produce recombinant proteins with proper post-translational modifications and folding, which are important for biological activity. In addition, mammalian expression system doesn’t have endotoxin problem in E.coli expression system. Commonly used cell lines include human 293 cells and CHO.
Bon Opus has decades of experience in mammalian cell expression. Our in-house protein experts will help you explore different options and work closely with you each step of the way.
Feature Package: Pilot-scale Mammalian Protein Expression ($950)
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Include gene synthesis, cloning, transfection-grade DNA preparation
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Transfection and expression in 20ml cell culture
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One-step purification with tag (His or Fc)
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6-8 weeks delivery time
General Workflow
1. Gene synthesis
2. Cloning & Transfection
3. Expression
4. Purification
Steps Breakdown
Signal Peptide
Signal peptide is critical for secretion in mammalian cells. Based on the protein of interest, we will help determine the best signal peptide to use. The options include common signal peptide such as IL-2, CD33 leader sequence, as well as our in-house developed signal peptide sequences.
Fusion Tags
The addition and position of epitope tags will be decided by the application of the protein. Commonly used tags include His, FLAG, and Fc.
Gene Synthesis & Cloning
Once a protein of interest is identified, the AA will be subjected to codon optimization to increase translational efficiency. The optimized gene will be synthesized and sub-cloned into our in-house developed expression vector. The entire process can be completed within 2 weeks.
Pilot Evaluation
Once an expression vector is made, a trial transfection will be conducted to determine the rough yield of the protein. The purified samples from the pilot can also be used to test its bioactivity in POC experiment.
Scale-up Production
Scale-up transfection will be performed according to the predicted yield and quantity need. It normally takes 2-3 weeks to complete. Based on the epitope tags and purity demand, affinity purification (protein A and his) along with other purification methods (IEX, gel-filtration, HIC, etc.) will be carried out to purify the target protein.
Final QC
After all procedures are completed, the proteins will be subjected to a round of final QC, which can include SDS-PAGE (reducing and non-reducing), concentration measurement (BCA, Bradford or A280), western blot, SEC-HPLC, endotoxin removal lowest to 0.1EU/mg.
Bon Opus staffs a team of industry-leading experts with decades of field experience and research dedication under their belts. We work closely with unique reagent variations to best serve our community of clients. Our commitment to understanding and providing mammalian cell protein expression services fosters the knowledge and innovation necessary to receive quality protein partnerships. We established our tailored services based on our ability to curate affordable, flexible, proactive approaches, ensuring each clientele feels understood from start to finish. We take you along our comprehensive approaches, beginning with gene synthesis and progressing through the cloning and transformation stage, into the expression development process, and through the finalization of all productions with an in-depth purification process. Our recombinant protein production in mammalian cells bridges the gaps in primary biological activities between folding and post-translational modifications.
Bon Opus honors the time needed to produce and provide reliable expressions. We achieve a 92 percent success and satisfaction rate with a six- to eight-week turnaround for our mammalian proteins. Our custom services create space for collaboration and detail. Fill out the easy online contact form to learn more.