Feasibility study.
We are protein biodesigners
Each new protein is an individual project and an exciting new challenge for our team. To succeed, this protein production project must be effectively designed from the onset.
We first follow a rigorous process, a feasibility study, to ensure that our customers are fully satisfied with their protein of interest.
Feasibility study: key stages
1/ Discussion of your project and expectations
2/ Preliminary study and quotation
3/ Option - In silico engineering
4/ Development of the expression vector(s)
5/ Screening of expression and purification conditions
6/ Feasibility study and dispatch of sample(s)
Discussion of your project and your expectations
As soon as you have informed us of your request, we will set up an initial telephone or video meeting to discuss your project, in order to fully understand your needs and objectives. A confidentiality agreement can be put in place, if necessary at this stage.
Preliminary study and quotation
Literature review and sequence analysis
Based on the information given by the client, we will search the literature and/or analyze the articles provided to identify the necessary conditions and the system that will work best to express your protein of interest. We will then gather all the conditions that have a positive impact on the expression of the protein as well as assess any potential difficulties that we may encounter.
Using protein databases and our sequence analysis tools, we will establish, based on this preliminary study, a work plan, and an estimate of costs to quickly and efficiently evaluate our ability to produce your protein of interest.
Optional: in-silico protein engineering
Synthelis has developed extensive partnerships with artificial intelligence companies which have developed proprietary solutions to predict in-silico the characteristics of the proteins. By leveraging these relationships, we can generate very quickly, and effectively, a library of mutants that will have the specific or optimized property you are looking for in your product or reagent: increased stability, improved activity, stronger affinity, better specificity etc.
Design and construction of the expression vector
The design of the DNA construct is also a critical stage in the process; one where the experience of a specialist plays a key role and increases the chances of success for a protein production project.
The right choice of expression tag (6xHis, Avi, Strep etc.), its positioning in relation to the gene, the size of the sequence between the tag and the coding region, the selection of the promoter etc, are all factors influencing the level of expression, the purification and the correct folding of the protein.
With more than 11 years of experience in protein production and a network of partners specialized in molecular biology, we can identify the DNA construct(s) that will enable the optimal expression and purification of your protein of interest.
Screening of expression conditions
Once we have a ready-to-use DNA construct, we will test the different key parameters for each expression system used:
– Salt concentration, cell extract type, T°C etc. for the cell-free protein synthesis system (CFPS).
– T°C, medium, clones etc, for E. coli based systems
The combination of parameters giving the best expression level and a whole protein, soluble if necessary, will be selected at this stage by Western Blot analysis.
– Estimation of the expression yield.
When the best expression conditions are defined, a small-scale production is carried out to allow the development of the purification protocol and to estimate the production yield of the protein. The resulting purified product is then either shipped to the customer for testing or used for quality control testing at Synthelis.
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