E. coli Expression
ProMab Biotechnologies has years of experience providing CRO services with a skilled team of scientists who will work with you in developing your protein or peptide of interest. We offer protein expression services utilizing e. coli expression, baculovirus expression, mammalian expression, gene synthesis and peptide synthesis, as well.
Our high-level recombinant protein E. coli expression service includes scale-up as requested, tag removal as requested, fast turn-around time, and competitive pricing. E. coli expression is among the most widely used systems for recombinant proteins because of its many advantages.
- Inexpensive setup and running costs
- High recombinant protein production levels
- Short timeline from cloning to protein recovery
- Limited technical knowledge required for culturing
- Scalability from small (1 mL) to very large culture (>10,000 L) volumes
E. coli expression takes 6 to 8 weeks from subcloning to expression in 2 phases starting at $3,200.
Subcloning into Expression Vector
- Amplification/isolation of the gene of interest (GOI) from a client-supplied construct and subcloning into a suitable transfer vector with His-Tag (or GST-tag).
- Sequence confirmation of insert.
- Mini-induction to over-express the target protein.
- Test for recombinant protein by SDS-PAGE and Western Blot. The customer must provide appropriate antibodies to the protein of interest if desired.
Estimated time: 3-5 weeks
Expansion and Purification
- One liter of bacterial culture will be induced with IPTG and harvested for protein purification.
- Ni-NTA purification of protein with 6X His-tags or glutathione for GST-tagged proteins using beads either under native or denatured conditions.
Estimated time: 2 weeks
- 5 μg of transfer vector
- Recombinant Protein
- Yield ranges from 0.5 – 50 mg/L of culture with an average yield of 3 mg/L. Solubility is not guaranteed.