Protein Engineering
Genebeyond® protein engineering services include:
- Protein (fusion protein) design
- Plasmid and expression vector design
- Multiple protein expression platforms (E. coli, yeast, baculovirus/insect cells, mammalian cells, cell-free protein synthesis)
- Protein and protein complex expression and co-expression
- Animal cell culture and fermentation process development
- Protein purification process development and scale-up
- Protein analysis and functional characterization
- Monoclonal antibody (mAb) and bispecific antibody (bsAb) design and production
- Unnatural amino acid-containing protein design and production
- Protein conjugation
Genebeyond® provides multiple proprietary protein expression platforms to meet various demands of different unique protein. Multiple E. coli expression systems can obtain the protein in 1-2 weeks at low cost and high yield. Our yeast expression system can provide high-yield bulk eukaryotic proteins with low cost culture condition. Our mammalian cell expression systems (CHO, HEK293, Hybridoma) offer active proteins with high-quality, especially requiring posttranslational modifications for the full or better functions such as IgG, GPCRs, cell/nuclear receptors, transcription factor, etc. Flexible options of transient expression and stable expression technology are provided for different projects or different research phases. Genebeyond also provides baculovirus/insect cell expression platform.
Genebeyond also offers state-of-the-art protein purification technology including affinity chromatography (AC), IMAC, ion exchange (IEX), gel filtration (SEC), hydrophobic interaction (HIC), and combination, process integration, optimization and scale-up.
1. Recombinant protein expression
Gene codon optimization
Plasmid design and construction
E. coli
Yeast
Baculovirus/ Insect cells
Mammalian cells (transient or stable expression)
Transient transfection and expression
Stable single cell clone and protein production
Cell-free protein production
Fusion protein
Difficult protein expression
2. Biopharmaceutical process development
Single cell line cloning
Fed-batch cell culture process optimization
Protein and antibody purification (AC, IEX, SEC, HIC, HPLC)
Upstream and downstream process integration and scale up
Comparison of protein expression systems
| Important Factors | Characteristics | E. coli | Yeast | Insect cells | Mammalian cells | Cell-Free (CFPS) |
| Functionality | Protein Folding | Refolding Usually Required | Refolding May Be Required | Proper Folding | Proper Folding | Tunable |
| N-linked Glycosylation | None | High Mannose | Simple, w/o Sialic Acid | Complex | None | |
| O-linked Glycosylation | No | Yes | Yes | Yes | None | |
| Phosphorylation | No | Yes | Yes | Yes | None | |
| Acetylation | No | Yes | Yes | Yes | None | |
| Acylation | No | Yes | Yes | Yes | None | |
| gamma-Carboxylation | No | No | No | Yes | None | |
| Solubility | Extracellular Expression | Inclusion body or soluble/secreted | Soluble/secreted | Soluble/secreted | Soluble/Secreted | In vitro |
| Success Rate (%) (soluble or functional) | 40-60 | 50-70 | 50-70 | 80-95 | Tunable | |
| Yield | Expression Level | High | Low – High | Low – High | Low – Moderate | High |
| Yield (mg/L ) | 0-1000 | 0-20,000 (22g/L) | 10-500 | 0.1-10,000 | 0- 2,000 | |
| Cost | Medium Cost | Low | Low | High | High | Low |
| Project Cost | Low | Low | Middle | High | High | |
| Speed | Cell Growth | Rapid (40Min) | Rapid (120 Min) | Slow (18-24 H) | Slow (24 H) | Rapid |
| Complexity | Growth Medium | Minimum | Minimum | Complex | Complex | None |