We provide small to large-scale protein expression and purification services in each of or 5 platforms at competitive prices but also more advanced and customized packages in which we test for instance custom recombinant protein expression conditions in several systems in parallel or with extended protein expression conditions in a given system. Our services include all the steps from gene synthesis and expression vector construction to pilot protein production in flasks and large-scale protein expression in fermenters and bioreactors.
Our High throughput protein expression condition screening is intended to overcome challenges related to solubility, purity, functionality, productivity, refolding or formulation. Such a commitment involves having access to, and mastering, all existing technologies, processes and tools available. With extensive experience in the expression of recombinant proteins of any kind, such as antibodies, enzymes, membrane proteins, cytokines, etc., and of any species, ProteoGenix has developed a complete toolbox to tackle any such challenge and to provide best-in-class protein engineering services. Try guaranteed packages from the most reliable protein production company of its kind.
Custom Protein Production
” I recently chose ProteoGenix to produce a recombinant protein in HEK293 cells. The high productivity and strong activity of the protein obtained, probably due to proper glycosylation and folding, allowed us to generate exciting results in functional studies and reach new milestones in our research project. The quality of the service provided, and the smooth and kind communication I had with the account manager in charge of our project, were paramount to this success. I’m happy to recommend ProteoGenix for recombinant protein productions. “
Why Choose ProteoGenix for your Recombinant Protein Production?
Why Choose ProteoGenix
More than 14 years of outstanding track record in the production of difficult-to-express proteins
5 available protein expression systems and complete toolbox to overcome any particular protein expression challenges
One-stop solution from gene to protein, process development and large scale production. Consequences: faster service, increased success rate, less administrative work.
IP-free strains and cell lines
Very small to large scale productions with 1 to 4000 L batch
Productivity of up to 800mg/L in transient expression and 5g/L in stable expression.
Example of insect cells production of a Beta-Glucosidase
We were asked by a customer to produce a recombinant Beta-Glucosidase protein using a baculovirus expression system (Insect cell production). Our customer ordered a full Insect cell package, including small-scale expression and purification tests followed by a 1L pilot production and purification. We reached a yield of 370 mg/L and delivered 60 mg after small-scale expression. 1L pilot production was no longer useful and was therefore not charged to our customer. The services carried out included:
The cDNA coding for Beta-Glucosidase protein was chemically synthesized after sequence optimization for insect cells expression. It was then subcloned in a proprietary expression vector. A sequence coding for a 6His tag was added for further purification.
P1 VIRUS STOCK GENERATION
Figure 1. Analysis of insect cell expression during P1 generation.
Left. Coomassie Blue Staining. Right. WB with anti-His antibody (ECL substrate) “-“. Un-transfected
control culture. 1 and 2 are transfections with 2 different Bacmid clones
Conclusion of P1 virus generation step
A protein band matching with Beta-Glucosidase protein molecular weight is observed by SDS-PAGE in native protein extracts. This result is confirmed by Western blot (Figure 1, green arrows). Recombinant bacmid clone No1 was used for expression tests.
VIRUS AMPLIFICATION AND EXPRESSION TESTS
Figure 2. Protein Expression Tests with P2 stock.
Coomassie Blue Staining. Left. Cell type 1.Right. Cell type 2. “-“. Negative control culture.
Conclusion of expression tests
Expression tests with P2 confirm that the target is present at high level in native protein extracts (Figure 2, green arrows). Optimal conditions for native expression: Figure 2, red arrow = loi
Figure 3. Protein small-scale purification test. Coomassie Blue Staining.
Left. Purification profile. Right. Final QC of purification pool
IN. Input. FT. Flow through. W1-W3. Washing steps. E1-E9. Eluted fractions.
Conclusion of small-scale purification tests
Recombinant Beta-Glucosidase protein can be produced and purified in native conditions. The purity is ≥ 95%, and the production/purification yield is approximately 370mg/L. Baculovirus expression system was a perfect choice for this protein.
Which System choose for your Recombinant Protein Production?
COMPARISON BETWEEN 5 EXPRESSION SYSTEMS PROVIDED BY PROTEOGENIX -
PROKARYOTIC VS EUKARYOTIC CELLS EXPRESSION SYSTEMS.
From a general standpoint, the table below gives an idea of the advantages of each five different heterologous protein expression systems for production of recombinant proteins. Of course, features can vary from one protein to another. The more arrows there are, the better it is.
|Time||Cost||PTM||Low Endotoxin levels||Simple process scale up||Secretion|
|Yeast (P. pastoris,
The table below gives an idea of the advantages of each five different heterologous protein expression systems for production of recombinant proteins from a general standpoint. Of course features can vary from one protein to another. The more arrows there are, the better it is.
|Time||Cost||PTM||Low Endotoxin levels|
|Yeast (P. pastoris,
|Simple process scale up||Secretion|
|Yeast (P. pastoris,
TIME DURATION AND PRICE
|Expression system||Time duration||Price|
|E. coli||3-6 weeks||Starting from
|B. subtilis||4-6 weeks|
|Yeast (P. pastoris,
|Mammalian cells||7-9 weeks|
Small scale expression and purification evaluation with many expression conditions tested.
1L production and purification
NON EXHAUSTIVE MULTI-EXPRESSION SYSTEM PACKAGES
|E. coli / Yeast / Baculovirus /
|B. subtilis/ Yeast / Baculovirus|
|E. coli / Baculovirus|
The most commonly used system of recombinant protein purification is E. coli. Indeed, it is the simplest, fastest and most cost effective system for custom expression of a protein. Bacterial cultures grow much faster than insect or mammalian cells or yeast. It is therefore easier and quicker to test a large number of different conditions. More solutions are also available for testing and therefore allow a higher success rate in expressing recombinant proteins. Transient expression yields of recombinant protein are also generally much higher than in yeast, baculovirus / insect cells or mammalian cells. Production of recombinant proteins in E. coli (Escherichia Coli) is often the preferred solution proposed by ProteoGenix when the final application requires large quantities of proteins such as crystallography and nuclear magnetic resonance (NMR) and no Post-Translational Modifications (PTM) are required. However, the main drawbacks of E. coli system are the high amounts of endotoxins remaining in the final sample and the lack of Post-Translational modifications. It is possible to remove endotoxins but it isn’t convenient at an industrial stage when there is not much to be done to solve PTM problems apart from using a different expression system. With these five systems, we are able to meet the requirements of a large number protein synthesis orders and we therefore have a success rate of over 95% in recombinant protein synthesis. We are constantly working on integrating new systems and solutions to provide the best level of service for anyone who needs to buy high quality recombinant protein production services.
Team, Capacity, Organization
If you have any doubts about which system to choose from, we, as a leading protein production manufacturer, can guide you from gene to protein and help you define an optimal strategy while taking into account your time constraints, budget and required protein features..
Our recombinant protein production laboratories are organized across several platforms and supporting labs to ensure the best quality of service, the highest product purity, the lowest risk of contamination and the best organization for a fast and reliable service.
We have a platform for each of our five systems and a separate quality control lab, molecular biology lab, packing lab… Each platform is managed by experienced team leaders that ensure they deliver proteins corresponding to initial customer specifications. With more than 1500 proteins produced to date, and specialized teams for each system, our expertise is unrivalled.
Custom recombinant protein
ProteoGenix recombinant proteins are used in a wide spectrum of applications and our customers are usually involved in Life sciences, including Pharma companies, Diagnostics, Biotech, the Agri-Food Industry, Crop science, New energies, Cosmetics, National Research Laboratories, etc..
Below is a non-exhaustive list of applications: