RD-Biotech offers a large range of tailor-made services on its recombinant protein platform.By contacting RD-Biotech you will:
RD-Biotech offers standard services but also tailor-made strategies adapted to your specifications.
Our experts are at your disposal in order to define the best strategies, to optimize the development of your project, in terms of time and cost.
Recombinant protein expression platform:
A MULTI SYSTEM OFFER
Expert service and in silico feasibility study
Analysis of sequence configuration, protein structure, predisposition analysis of protein expression, structural biology (Spectrometry, Dichroism, Biacore ...)
Construction of the expression vector
In case you do not have the expression vector, we offer the design and cloning of the suitable and optimized vector for the desired expression system.
Feasibility study
Pilot expression and purification, tests for different expression conditions (evaluation of different strains or cell lines in parallel, tests for different culture media, temperature conditions, etc.): determination of optimal performance conditions and expression performance, analysis of the results and report.
Customer GO / NO GO for large scale productio
Production and purification of the desired quantity
Quality controls
- Standard controls: SDS PAGE, agarose gel electrophoresis, spectrophotometry
- Additional controls according to request: ELISA, Western blotting, Cytometry, Endotoxins, Host Cell Proteins, activity tests ...
Supplying of the batch of purified protein
accompanied by the production / purification report and quality controls (buffer, concentration, aliquoting)
THE RD-BIOTECH QUALITY
- Optimized production systems adapted to your applications: bacterial, mammalian, yeast and cell-free systems.
- “Multi-system expression” offer: Possibility to test different expression systems in parallel.
- Molecular biology platform for a complete service, from the construction of the expression vector to the supply of the purified and controlled protein
- R&D quality and “endofree” quality
- Capacity: production of pilot batches and large-scale production (up to several hundred mg depending on the system)
- Speed: Supply of purified and controlled batches in less than 3 weeks.
Bioproduction
Expression of recombinant proteins in E. coli system
- An expertise of more than 20 years in bacteriology, with the expert support of the mAbexperts group
- This prokaryotic system offers the best compromise between expression yields and production costs.
- A set of tests, complementary to the standard process, to optimize the yield and quality of the proteins produced and purified
Scale-up production capacity: from a few mg to several hundred mg
- (the vector can also be provided by the client)
- Synthesis of the gene of interest (if necessary)
- Subcloning in the expression vector. Screening
- Sequencing control
- Amplification and quality controls
- Pilot study based on culture volumes of a few dozen ml
- Protein control induced by SDS-PAGE gel electrophoresis
- Western-blotting control (anti-tag antibody or specific antibody)
- Protein purification by affinity chromatography on Nickel Agarose (HIS tag) (other method possible, contact us)
- Quality control: Control of the purified protein by electrophoresis on SDS-PAGE gel and protein assay
If necessary: Additional expression tests:
- Transformation of different bacterial strains
- Selection of the best producers
- Tests of different expression conditions (induction type, temperature …)
- Renaturation tests
- Solubilization tests
Go No Go
Scale-up production capacity: for Research batches (batches from a few mg up to several hundred mg)
- Higher volume production
- Purification by affinity chromatography on Nickel Agarose (or other method)
- Control of the purified protein by electrophoresis on SDS-PAGE gel and protein assay
- Option: Elimination of the tag
- Possibility of concentration, aliquoting, lyophilization, conjugation …
- Packaging and dispatch of the protein, transport in dry ice, accompanied by the report
Expression of recombinant proteins in mammalian cells: CHO, HEK293,...
- Broad expertise on many cell lines: adherent cells and cells in suspension
- System allowing conservation of the native structure and post-translational modifications
- System for simple and rapid evaluation of protein expression by transient transfection
- Pilot study in culture flasks or plates on small culture volumes
- Control of expressed proteins: Western-Blotting, ELISA, flow cytometry …
- Protein purification: different systems available, depending on the protein (purification by affinity chromatography, etc.)
- Western-blotting control (anti-tag antibody or specific antibody) and SDS-PAGE gel
Scale-up production capacity:
- from a few ml to several hundred litres of supernatant per batch
- from a few million to several billion cells per batch
- Synthesis of the nucleotide sequence of interest (if necessary)
- Subcloning in the expression vector optimized for the cellular system
- Sequencing control
- Amplification and quality controls.
- Pilot expression: Transfection, production monitoring of the recombinant protein.
- Pilot purification by affinity chromatography. (Option: purification under “Endotoxin – free” conditions).
- Quality controls:
- Purity control by SDS-PAGE electrophoresis (reducing and non-reducing conditions).
- Quantification
- Others depending on the project: Western blot, …
- Packaging and dispatch of the recombinant protein accompanied by the production certificate.
Go No Go
Expression of recombinant proteins in Cell Free in vitro system (WGE)
The advantages of this system:
- Cell-free process
- Excellent alternative for the expression of proteins toxic to cells
- Expression of soluble or membrane proteins
- Better protein folding
- No codon optimization step
- Wide temperature range: 4 to 30 ° C
- Open system: possibility of adding detergents, phospholipids, markers …
- Vector optimized for this cell-free system
- Production capacity up to several mg
Hundreds of constructions carried out each year (membrane proteins, viral proteins, membrane receptors, GPCR like proteins,…)
- Synthesis of the nucleotide sequence of interest (if necessary)
- Subcloning into the expression vector optimized for the cell free system.
- Sequencing control
- Amplification and quality controls.
- Pilot production and purification
- Control of proteins induced on SDS-PAGE gel
- Western-blotting control (anti-tag antibody or specific antibody)
- Protein dosage
If necessary: Additional expression tests:
- Tests of different expression conditions
- Solubilization tests
Go No Go
Expression of recombinant proteins in yeast system (P. pastoris, P. cerevisae)
Why choose yeast system:
- Post-translational system similar to that of mammalian cells, including glycosylation, phosphorylation, …
- Relevant system for complex proteins
- Secretion into the culture medium
- “Endofree” quality
- High production yield
- A set of tests, complementary to the standard process, to optimize the yield and quality of the produced and purified proteins
Scale-up production capacity: from a few mg to several hundred mg.
(the vector can also be provided by the client)
- Synthesis of the gene of interest (if necessary)
- Subcloning in the expression vector. Screening
- Sequencing control
- Amplification and quality controls
- Test of different conditions of expression (e.g. induction T°C,…).
- Protein purification by affinity chromatography on Nickel Agarose (HIS tag)
- Quality control: Control of the purified protein by electrophoresis on SDS-PAGE gel and protein assay
Go No Go
Scale-up production capacity: for Research batches (batches from a few mg up to several hundred mg)
- Higher volume production
- Protein purification by affinity chromatography on Nickel Agarose (HIS tag)
- Quality control: Control of the purified protein by electrophoresis on SDS-PAGE gel and protein assay
- Option: Elimination of the tag
- Possibility of concentration, aliquoting, lyophilization, conjugation…
- Packaging and dispatch of the protein, transport in dry ice, accompanied by the report
Comparison between the 4 expression systems provided by RD-Biotech
The table below gives a general idea of the advantages of the three heterologous protein expression systems available from RD-Biotech. However, the performance shown may vary from protein to protein..
Custom services
In silico feasibility studies
Expertise service and in silico feasibility study: project support including a scientific and technical analysis of the bibliography, study of sequence configuration, analysis of the predisposition of protein expression, structural biology, in order to propose the most appropriate sequence and system.
Vector construction
RD-Biotech offers two alternatives:
- Using your vectors
- Development of your expression vectors in our optimized systems
Other services
Our sales team as well as our scientific and technical experts are at your disposal to offer you the service adapted to your request!