Workflow of Bacterial Protein Expression (E.coli)
Process of Recombinant Protein Expression in E.coli
Quote and Project Plan
After your (inquiry), we discuss your individual project requirements and our scientific experts run an initial (feasibility check) with your protein sequence to define the ideal service strategy for this specific target.
You will receive a detailed project quotation by email or through your preferred secure data transfer method. Once the order is confirmed, the project commences within one business day. Our dedicated project management team will provide continuous updates to ensure optimal progress and successful outcomes.
Template DNA
The cloning workflow starts after receiving the template DNA. This can be supplied by the client as plasmid DNA containing the codon‑optimized sequence of the target gene. Alternatively, trenzyme offers gene synthesis of sequence‑optimized DNA as an optional add‑on, ensuring optimal compatibility with the expression system.
Plasmid Generation & Cloning
After we receive the template DNA, the following steps are:
- PCR-based cloning or direct cloning of target DNA into trenzyme´s proprietary expression vector(s) from our toolbox of more than 400 validated plasmids.
- Isolation of plasmid DNA
- QC of final construct by DNA sequencing
Small Scale Pilot Study
To enhance efficiency and reduce both time and cost for our clients, we conduct a preliminary pilot study on a smaller scale. This initial evaluation allows us to assess and refine, ahead of full-scale production, how the target protein can be effectively expressed within our available systems and whether the purification process is technically feasible. For more details on what this pilot study entails, see our (General FAQs).
Production Step 1: Protein Expression
The expression of your target protein is carried out in stirred flasks or shake flask culture based on trenzyme's optimized standard protocol and the conditions of the specific target protein determined in the (pilot study).
Production Step 2: Protein Purification
After successful expression, the protein is purified. For the expression in E.coli , this includes:
- One-step purification of target protein from culture pellet at 1 ml affinity resin scale according to data from the pilot study
- Buffer exchange into standard buffer of your choice
Final Quality Control of Protein
Before we send you your purified target protein, a final quality check is performed by:
- Analysis by SDS-PAGE
- Assessment of final concentration and purity
Shipment of Your Protein
- Global priority shipping
- Quick and efficient delivery under optimal conditions for your protein
- Trusted international logistics partners
- Real-time shipment tracking for full transparency
For further details, see our (FAQ)s.
Deliverables
Together with your purified target protein you will obtain a datasheet. Which data we provide is detailed in our (FAQs)
Timeline
The complete workflow of production of the target protein in E.coli is usually realized in 4-6 weeks.
What Our Clients Say About Us
“(…) trenzyme has been a reliable, flexible, and innovative outsourcing partner. (…)”
Recombinant Protein Expression in E.coli – Service Package for Standard Protein Targets Includes
Additional Service Options:
trenzyme’s E. coli Recombinant Protein Production Service – Your Benefits
Guarantee – Pay Only When We Succeed!
After feasibility analysis and successful pilot study we guarantee the final quantity and purity of your desired target protein in the given timeframe. You only pay when we succeed!
Comprehensive Service Package
Comprehensive service package for recombinant protein expression and purification for a fixed price and a fixed scale.
Transparency & High Quality
Our dedicated project management will guide your project through every step of the process and keep you constantly informed on the progress. Transparency and high quality service are our mission.
Free Feasibility Analysis & Consulting
Our free feasibility check of your protein sequence is the basis for our free individual consultation with you to define the ideal service strategy for your target that will lead to success.
Standard Protein or Challenging Target – Evaluated by Feasibility Check and Pilot Study
During initial free individual consulting, our scientific experts run a feasibility check (e.g. literature check, database check, product check, etc.) with your protein sequence to define the ideal service strategy for your specific target.
To achieve maximum efficiency and save you significant time and money, we additionally run a small scale pilot study in this service process. These preliminary assessments help us determine in advance whether the target protein expression in E.coli is possible and whether the purification process is feasible. If desired, we will provide you with test samples to check the functionality and quality of the purified target protein in your hands.
The big benefit for you: After pilot scale evaluation we guarantee the final yield, purity and timeline – you only pay the production run when we succeed!
Recombinant Protein Expression Service in E.coli
Application Note: Recombinant expression of 15N-labeled protein in E.coli by high cell density cultivation
Reinhold Horlacher, PhD
Managing Director & CSOWe would be happy to provide you with support on your protein research project.
Get In Contact With Our Scientific Experts
Design Your Protein Now!
Use our user-friendly protein configurator to create your individual protein according to your needs. Click on the button below to get started!
Questions or Individual Quote?
Let us know your questions and requests by simply filling out the following contact form. Our scientific experts will get in touch with you immediately to discuss your request.
