Recombinant Protein Expression in E.coli 

Quote and Project Plan

• After your individual project requirements have been discussed, our scientific experts run an initial feasibility check with your protein sequence (e.g. literature check, database check, product check, etc.) to define the ideal service strategy for this specific target.
• The detailed quote for the project is sent to you via e-mail or by secure data transfer of choice.
• The project will start within one working day after order confirmation – from this point of time our dedicated project management team keeps you constantly informed throughout the project to ensure maximum success.

Template DNA

• Template DNA delivered from client: plasmid DNA containing the codon optimized DNA of the POI (Protein of interest)
• Easy alternative - Gene Synthesis Service by trenzyme: Gene synthesis of codon and DNA structure optimized DNA (available as add-on).

Plasmid Generation & Cloning

• PCR based cloning or direct cloning of target DNA into trenzymes’ proprietary expression vector(s)
• Isolation of plasmid DNA
• QC of final construct by DNA sequencing

Small Scale Pilot Study

To achieve maximum efficiency and save the customer significant time and money, we run a small scale pilot study. 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. For validation, we follow the steps outlined below:

• Transformation of construct into E.coli expression strain
• Generation of cryostock
• Small-scale protein expression in culture
• Small-scale purification by cell lysis and affinity chromatography (batch mode)
• Validation of successful target protein purification by SDS-PAGE and Western Blot analysis
• Compilation of documentation of pilot study
• Guarantee: After a successfully completed pilot study, the outcome of this service package (purity and yield) is guaranteed
• Flexibility and Efficiency: If expression and purification is not feasible as desired, the project can be stopped at this point. Therefore, no further costs and time are incurred. However, we won't leave you alone with your demanding target - our ➥Custom Protein Production Service provides a huge variety of highly customized solutions in order to achieve the project goal under modified conditions.

Production: Protein Expression

Protein expression in shake flask culture according to trenzyme's optimized standard protocol (up to 4 L cultivation volume - Higher yield possible via ➥Custom Protein Production Service ).

Production: Protein Purification

• One-step purification of soluble target protein from culture pellet at 1 ml affinity resin scale according to data from the pilot study
• Buffer exchange into standard buffer

Final guaranteed yield: 1 mg and purity: 90%

Final Quality Control of Protein

• Analysis by SDS-PAGE
• Assessment of final concentration and purity
• Compilation of comprehensive datasheet

In case we are able to produce more than the guaranteed final amount of target protein, the excess protein can be purchased.

Shipment of Your Protein

• Fast and reliable worldwide delivery
• Our logistics specialists and trusted shipping partners guarantee that your products reach you in excellent condition and without any loss of quality
• Collaboration with established global courier services operating an extensive logistics network
• Full shipment transparency through continuous parcel tracking; you will receive the tracking code once your order is dispatched
• Proteins and cell lines are typically shipped on dry ice to maintain stability and quality. Upon request, we can also arrange alternative shipping options such as dry shippers, blue ice, or ambient temperature transport
• We handle every delivery responsibly, as environmental sustainability and climate protection are central values of our company (see ➥ Code of Conduct).

Deliverables

Together with your cryopreserved cells you will obtain comprehensive documentation including

• Data sheet with exact culture conditions and media compositions
• Growth rate documentation
• Optional: a full report about the whole adaption process

Timeline

The timeline very much depends on the cell line, the doubling rate of the cells and on the type of medium (low or high nutrient content) to which the cells shall be adapted. Therefore, timeline ranges from 6 to 12 weeks.