Recombinant Protein Production

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.

Recombinant Protein Production

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