Recombinant Protein Production
We develop strategies in E. coli, Insect and mammalian expression systems to produce proteins specific for your application. Our team can execute both technical transfer of your protocol or develop a customized process.
Expression in Mammalian Cells
| Service Options | Details |
|---|---|
| Cell Lines | HEK293, CHO-S, HEK293-6E, CHO-3E7 (EBNA), others by request Adherent cell lines and others upon request |
| Transfection | PEI, Lipid-based reagents |
| Expression Test and Optimization | Cell lines, plasmid ratios, harvest time Western blot on cell extracts or media with optional small-scale affinity purification |
| Expression, Flask | Up to 45 L currently, readily expandable up to 36 X 0.5 L to 2 L scale in parallel for antibody production up to 90 x 50 mL for antibody screening |
| Expression, Bioreactor | 2 X 25 L (System 20/50 WAVE Bioreactor™) |
Expression in Insect Cells
| Service Options | Details |
|---|---|
| Cell Lines | Sf9 and Sf21 standard, others by request |
| BV Generation & Amplification | Homologous recombination or transposition based |
| BV Titer Count | Viral plaque assay or alternatively BIICS |
| BV Expression Test and Optimization | Western blot cell extracts or media, 6 - 18 data points per construct Test MOI, TOI, cell lines Monitor infection by cell diameter, cell viability and cell count |
| BV Expression, Flask | Up to 18 L currently, readily expandable |
| BV Expression, Bioreactor | 2 x 25 L (System 20/50 WAVE Bioreactor™) |
| Transient Expression | Plasmid mediated, expression screen and scale-up |
Expression in Bacterial Cells
| Service Options | Details |
|---|---|
| E. coli Strains | Strains selected for each project, all common expression strains are available |
| Expression Optimization | SDS-PAGE or WB extracts (total, soluble and insoluble protein) multiple constructs, strains, media, and induction temperatures, 6 and 12 data points standard, optional micro affinity purification |
| E. coli Expression, Flask | 60L per expression (15° - 40° C) and 5 runs per week |
| E. coli Expression, Fermenter | 10 - 440 L |
| E. coli Expression, Fermenter | Custom fermentation development 4 X 5 L or 8 X 1 L in parallel |
Protein Purification
| Service Options | Details |
|---|---|
| Tech Transfer or Development | Reproduce client methods or develop de novo strategy |
| Chromatography | All common techniques including affinity, IEX, HIC, SEC AVANT 25 & 150 and AKTA Explorers |
| Scouting | Develop or refine methods by scouting >10 or more conditions per batch |
| Scale, Throughput | Microgram scale up to 50 g. Parallel purification options available for HT. Up to 90 samples per lot in batch mode, up to 36 samples per lot using column mode purification |
| Refolding, Dilution or Dialysis | Custom 24 condition dilution refolding screen up to 200 L scale dilution refold |
| Formulation Screen/Stability Study | 96 condition formulation screen (buffer/excipient/detergent/salt) or custom. Monitored by turbidity, concentration, SEC-HPLC, target-specific activity assay. Data can be used for purification method development and long-term storage |
Quality Control
| Service Options | Details |
|---|---|
| PAGE | Denaturing, reducing, non-reducing, densitometry (Bio-Rad ChemiDoc XRS+) |
| Concentration | Dye binding assay, A280, RP-HPLC |
| Western Blot | Anti-Tag or Anti-Target, Colorimetric or chemiluminescence (ChemiDoc XRS+) |
| Analytical SEC | Standard or client-specific methods |
| Reverse Phase HPLC | Standard or client-specific methods, purity and / or concentration measurement |
| Mass Spec | Whole protein mass by MALDI or ESI, in-gel digest, LC-MS/MS, by qualified partner |
| Endotoxin | LAL Assay |
| Activity Assay | Absorbance, fluorescence, luminescence (Cuvette to 384-well plate) |
| ELISA | Anti-tag or anti-target, host cell protein, commercial or custom |
