Custom Designed Recombinant Proteins

Custom Designed Recombinant Proteins

In recent decade, genomic and proteomic tools has enabled to identify genes and proteins potentially involved in different biological process.

However, the characterization of a part of theses proteins is hampered by the availability and cost of a pure and active protein. Tailor-made recombinant is time consuming, requires an expensive facilities and an extensive know-how related to genes and proteins handling (cloning, cell culture and purification).

PROTENIA is a CRO, plainly devoted to the recombinant protein production and engineering.

For each project, we study the feasibility; optimise the expression level and protein yield before scaling-up the production. You can entrust us with your project at different stages :

• from gene to protein: genes are either cloned from a cDNA provided by client. Otherwise, we synthesize the gene (codon optimisation, directed mutagenesis),
• from a provided construct to protein : we achieve the expression and optimisation tests, purification and yield evaluation.

Gene Isolation & Synthesis

Gene Isolation & Synthesis

We can isolate for you the gene of interest either by RT-PCR or PCR from provided mRNA, cDNA or DNA. We also have the gene synthesis service for codon optimization, mutagenesis, etc. This step is validated by sequencing.

Cloning/Subcloning in Expression Vectors

Cloning/Subcloning in Expression Vectors

We clone routinely into a wide range of vectors for expression in E. coli, Yeast, and insect cells carrying different tags and regulatory sequences. For special requirement, vectors can be customized according to your needs (Tag, promoter, …). Each construction is validated by sequencing and reported on.

Protein Production scale

Protein Production scale

In all cases, for functional and biochemical characterization (enzymatic activity, binding…) and structural studies we provide amounts of protein ranging from µg to grams.

Once the production and purification parameters are optimized, or transferred when available elsewhere, we go to the final batch production:

• Lab-scale production (0,5-5L)
• Pilote scale: for a milligram/gram-scale production, a set of fermentors (1L, twin-5L, 30 L) enable us the convenient scaling-up.

Protein Purification

Protein Purification

This step is crucial for the success of the project. Our expertise and know-how associated with suitable facilities guarantee purification in the best conditions, saving thereby the integrity and properties of proteins.

Basically, the purification is based on affinity chromatography. However, if a high purity grade or polishing step is required, we combine/associate additional techniques: ions exchange, gel filtration, hydrophobic interaction chromatography. We also purify protein without tags.

You can entrust us your production projects off or regular:

• Recombinant enzymes
• Recombinant antigen (diagnostic kit, ELISA)
• Recombinant antibodies
• All recombinant proteins and peptides

The proteins are expressed in different prokaryotic systems (E. coli) and/or eukaryotic (yeast and insect cell / baculovirus, as cytosolic or secreted proteins).

Depending on your needs we can deliver proteins endotoxin free, and high levels of purity> 97% for structural studies (crystallography, NMR).

Delivery time

Delivery time

It depends on gene size, expression system, protein type and required purity. Here are the required time for each step and expression system.

Initially, we conduct a feasibility study for the project and set up the necessary protocols for protein expression and purification. After validation of the feasibility study (production assay, yield evaluation), a report is sent to the client.

The proteins obtained in phase A can be delivered if the customer wishes, particularly to achieve the first tests. After validation, we perform the scaling-up for the final lot asked.

However, if the results of the Phase A show that production of the protein is not possible under the selected strategies, the continuation of the project is then discussed with the client to stop the project or to change the strategy (vector, strain, expression system, Tag ...).