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Protein Sciences

Uniting Drug Development Expertise with Antibody & Protein Production Services

E. Coli Expression Systems

High-Yield E. coli Recombinant Protein Production

 

E. coli expression systems play a significant role in biologics development, especially for producing proteins where post-translational modifications (PTMs) aren’t crucial or can be addressed later in the process. Their speed, cost-effectiveness, and ability to yield large quantities make them invaluable tools in bioprocessing, particularly for research, early-stage development, and certain biologics manufacturing.

 

At WuXi Biologics, our Protein Sciences (PS) Department has extensive experience in E. coli recombinant protein production. We excel in scaling up E. coli fermentation, demonstrating expertise in producing both soluble proteins and refolding inclusion bodies. This enables us to consistently deliver high-quality recombinant proteins at scales ranging from small to large, catering to various research stages.

Scaling-up E. coli fermentation, E. coli soluble protein production, and inclusion body refolding expertise.

Key Features of Our E. coli Recombinant Protein Production Services

  • Comprehensive expression optimization steps to ensure high-titer expression (1-20 g/L)
  • Efficient scale-up from 1 L to 50 L
  • Customized services to meet specific requirement on purity and endotoxin level

Our expertise and strengths in E. coli recombinant protein production.

E. coli Recombinant Protein Production Service Details:

Service Item Deliverables Expression Scale Duration QC Request A Quote
Expression condition screening (4~8 conditions
per set)

Report in PPT format:

  • Host strain
  • Titer evaluation
  • Induction temperature and culture period
  • Induction OD
  • Medium
 5-10 mL

1 week

 SDS-PAGE Request A Quote
96DWP refolding condition screening Report in PPT format NA

2-3 weeks

 NA
Scale-up production in shake flask  Tagged or untagged protein, biotinylation    1-40 L 3-4 weeks   Concentration, SDS-PAGE (R/NR), and LC-MS
Endotoxin removal upon request
Scale-up production in bioreactor Tagged or untagged protein, biotinylation    1-50 L 4-5 weeks
Additional QC RP-HPLC, bioburden, DSF, DSC, SPR, LC-MS, SEC-MALS, peptide mapping, glycan profiling, cIEF, CHO HCP, residual DNA, ELISA, Western blot, freeze thaw study, micro developability testing

Case Studies: Optimizing Recombinant Protein Production in E. coli Expression Systems

 

The following three case studies demonstrate our expertise in E. coli recombinant protein production and the significance of expression optimization on yields and purity.

Case #1: E. coli Expression Optimization & Target Protein Expression

 

In this case study, systematic E. coli expression optimization substantially increased target protein yield and purity.

Figure A: Before expression optimization, the inclusion body showed very low expression, and in the pilot test, the protein was partially truncated after purification. Subsequent optimization efforts included experimenting with different hosts, culture media, and temperature conditions. As a result of these optimizations, the expression level increased by more than 20 times, and the protein achieved a purity of over 95% after the final purification step.

This case study shows that systematic E. coli expression optimization substantially increased target protein yield and purity for proteins with low expressions.

Case #2: Major Histocompatibility Complex (MHC) Protein Production from Small Scale to Large Scale Using E. coli Expression System

 

This case study showcases the successful production of complex MHCs across various scales through a robust refolding platform and precise purification.

Figure A & B: MHCs, which are ternary complexes composed of 3 crucial components, have presented challenges in terms of their expression and proper assembly, while maintaining their ability to present peptides effectively. These data showcase the successful production of 260 MHCs across a range of scales, from 0.1 L to 50 L. This accomplishment was achieved by employing a robust MHC refolding platform and a carefully designed purification strategy, which included in vitro biotinylation.

This case study showcases the successful production of complex major histocompatibility complexes (MHCs) across various scales through a robust refolding platform and precise purification.

This case study showcases the successful production of complex major histocompatibility complexes (MHCs) across various scales through a robust refolding platform and precise purification. width=

Case #3: E. coli Fermentation Production of an Enzyme for Bioconjugation

 

This case study underscores the importance of purification expertise in achieving a purity level exceeding 99% for an untagged enzyme, essential for bioconjugation.

Figure A: A 20 kDa untagged enzyme was expressed in a 50 L bioreactor. The most challenging part of this project was purifying the untagged target protein, a crucial step for its subsequent bioconjugation. Drawing upon our expertise in protein purification, we optimized the flocculation conditions by varying the concentration of PEI. Our purification process included CEX, HIC, and buffer exchange, resulting in the successful purification of the target protein to a purity level exceeding 99%.

o This case study underscores the importance of purification expertise (CEX-HIC) in achieving a purity level exceeding 99% for an untagged enzyme, essential for bioconjugation.

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