Mammalian Cell Expression System

The rapid progress of molecular cell biology has created a need for structural biologists to look at increasingly more complex protein structures. These usually require particular post-translational modifications and co-factors to support their elaborate folding and enzymatic activity. Prokaryotes are unable to perform these modifications and lack essential eukaryotic co-factors.

Mammalian cell expression exhibits the highest level of protein processing, which is important for complete biological activity. Therefore, the mammalian expression system is suitable for expressing complicated, modified, functional proteins/antibodies; secreted or membrane proteins, such as recombinant glycoproteins (vaccines and enzymes), hormones, antibodies, interleukins and lymphokines.

Owing to the low expression level of mammalian systems and the complex components of the medium, the expenses incurred for obtaining large amounts of protein are relatively high, which in turn poses a challenge in achieving large amounts of protein. With the top technical team and unique optimization scheme in BIC, we can promptly evaluate and identify the optimal expression conditions to meet your diverse needs.

BIC offers a 5% discount for all online inquiries! Contact us today and communicate with our scientists.

Highlights

• Rapid expression of functional protein or antibody.
• Customization: quantity, purity, endotoxin, and RNase level are optional along with any other services you may request.
• Modifications: isotope labeling, protein biotinylation, and green fluorescent labeling services.
• Our services advance your project from transient to stable expression.
• Expression scale-up: from 1 to 100 L

Mammalian Expression Service

Content	Service Process	Deliverables	Timeline
Gene synthesis	Codon optimization Gene synthesis Subcloning & Max-Prep	DNA sequencing data Cloning vector (1ug) Expression & purification results QC data	2-3 weeks
Transient Transfection Expression	Pilot Expression: Expression test Purification test Western blot analysis Scale Up: Protein/Antibody expression (0.2-1L) Purification: affinity chromatography, gel filtration, ion-exchange QC: SDS-PAGE & Western blot, mass spectrometry (optional)		4-6 weeks
Stable Transfection Expression	Stable Transfection Cell Line Construction

The Process of Protein or Antibody Production in Mammalian Cell

Additional Services

Based on our customized solutions and extensive experience, we provide the following two optional services:

Protein Labeling

Protein labeling technique refers to the covalent attachment of molecules such as enzymes to specific sites on the target protein. BIC has developed a series of protein labeling techniques, including fluorescent probes (FITC), enzyme conjugates (horseradish peroxidase and alkaline phosphatase), isotope labeling and biotinylation.

Large Scale Production

The WAVE bioreactor is the ideal cell culture equipment. WAVE utilizes a sterile and closed disposable bag, that can effectively prevent contamination. In addition, WAVE can automatically control process conditions such as incubation temperature, pH, dissolved oxygen, and pressure.

Case Study

Recombinant protein production in mammalian cell

Figure 1. 15kDa protein purification

Lane M: Protein marker; Lane 1: Culture supernatants of transfected mammalian cells; Lane 2: Protein purification, flow-through; Lane 3-5: Elution with different imidazole concentration buffers

Figure 2. Glycosylation modified protein expression

Lane M: Protein marker; Lane 1: Culture supernatants of transfected mammalian cells; Lane 2: Protein purification, flow-through; Lane 3-5: Elution with different imidazole concentration buffers

Recombinant antibody production in mammalian cell

Figure 3. Human IgG expression and purification

Lane M: Protein marker; Lane 1: Culture supernatants of transfected mammalian cells; Lane 2: Antibody purification, flow-through; Lane 3-5: Glycine elution

Figure 4. Fab antibody expression and purification

Lane M: Protein marker; Lane 1: Culture supernatants of transfected mammalian cells; Lane 2: Fab antibody purification, Flow-through; Lane 3: Glycine elution (Reduced); Lane 4: Glycine elution (Non-reduced)

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