Expression in mammalian cells

Mammalian cells are excellent hosts for the production of complex recombinant proteins that require extensive folding, the assembly of subunits and/or posttranslational modifications. For these reasons, mammalian cells are widely used by the biotech industry for the production of bio-active proteins.

Different mammalian cell types are used according to the quantity and quality of product required.

 

Service Description

All services include multiple steps from gene to purified protein as described in the table below.

Leadtime is approximately 10-14 weeks.

Description Resulting Material Average turnaround (*)
Gene Synthesis Stage
-Codon optimization
-Gene synthesis
-Cloning in standard pUC plasmid
-Complete sequencing
Cloned, synthesized and verified gene 1-2 weeks
Subcloning in Baculovirus Transfer Vector Stage
-Plasmid prep
-Subcloning Bac-donor vector
-Clone selection
-Complete sequencing
Plasmid vector with cloned target gene 2 weeks
Bacmid Generation Stage
-Plasmid prep
-Recombinant bacmid selection
-Recombinant bacmid DNA isolation
-DNA Verification
Bacmid DNA with cloned target gene 2 weeks
Insect Cells Transfection Stage
-Multiple transfection experiments
-Virus titering
-SDS-PAGE / Western blot analysis
High titer recombinant baculovirus stock 3 weeks
Pilot Expression Stage
-Expression pilot and optimization experiments
-Determine yields and protein behavior
10-50 µg of semi-purified protein 1-2 weeks
Expression and Purification Stage
-Expression
-Protein extraction
-Protein purification
-SDS-PAGE, WESTERN blot
0.1 to 1 mg of purified protein
Purity >95%
2-3 weeks

(*) Average production time for average difficult project. Actual turn around time varies with individual project features.

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