Protein expression optimization

Protein expression form

High throughput protein expression is the solution to overcome the most demanding challenges regarding difficult-to-express proteins! ProteoGenix developed a HTP platform dedicated to protein expression optimization. Our solution: 1000+ expression conditions carefully selected and tested in parallel to skyrocket your protein production in a short timeline.

Why choose ProteoGenix for
protein expression optimization?

protein expression conditions
Protein expression
“à la carte”

We design the most suitable expression conditions to test to meet the specifications

fast protein production
Save time

We can perform 1000 tests in 4 weeks Thanks to our fully automated high throughput platform.

high throughput protein production for difficult-to-express proteins
The ULTIMATE solution
for difficult-to-express proteins

A challenging protein expression? Our high throughput protein expression screening overcomes any challenge

protein expression expertise
Unique protein
expression expertise

We expressed over 1500 proteins since 2003.

protein expression systems
5 expression
systems

We have a deep understanding of 5 different expression systems: E. coli, B. subtilis, yeast, baculovirus/insect cells, and mammalian cells.

CHO protein expression
XtenCHOTM
inclusive!

Our service can include our high-performance mammalian expression system.

High throughput protein expression
for difficult-to-express proteins

ProteoGenix combines the knowledge of its best protein production experts to provide you the most advanced high throughput protein expression platform. Our high throughput platform was designed following one rule: “The more relevant conditions we test, the more probability we have to find the way to express your protein successfully”. Thus, our high throughput protein expression platform allows us to test 1000 conditions in 4 weeks in order to meet or even outperform your requirements in terms of:

  • Productivity

  • Purity

  • Solubility

  • Stability

  • Biological activity

Achieving such results is only possible by proposing a wide range of protein expression conditions. Here is a list of parameters ProteoGenix can vary:

MOLECULAR BIOLOGY CAPABILITIES
gene synthesis

Gene synthesis

UNLIMITED
vector for protein expression optimization

Vectors

UNLIMITED
signal peptide

Signal peptide

250+

Purification tag

15+
solubility tag

Solubility tag

15+
STRAINS AND CELLLINES
CULTURE CONDITIONS
temperature tests for protein expression optimization

Temperature

time tests for HTS expression

Induction
conditions

media for protein expression screening

Media
Screeining

extraction methods for protein production optimization

Extraction
Method

fermentation for high-throughput protein expression

Fermentation

UNLIMITED
PURIFICATION
purification of difficult-to-expression-proteins

Are you facing difficulties in protein expression optimization? Contact our PhD account managers to unlock the potential of your production.

Our high throughput
protein expression workflow

gene synthesis for protein production optimization
  • Determination of best host/vector pair

Test of several transformants
Selection of those presenting the best results
Purification tests on the best host/vector pair

GO / NO GO 1
Protein expression optimization in flask
  • Optimization of protein expression in flask

Optimization of several cultural conditions(temprature , induction time..)
Determination of the best feeding strategy

GO / NO GO 2
protein purification optimization
  • Optimization of protein purification
GO / NO GO 3
protein expression in bioreactor
  • Optimization of protein expression in flask

Optimization of fermentor/bioreactor
operating parameters
Protein purification of QC analysis: SDS/page,western

Example of a typical project

  • 4 E. coli strains
  • 2 gene synthesis
  • 4 mammalian cell lines
  • 4 vectors
  • 2 purification tags
  • 5 signal peptides
320
Tests

2 best system constructions combinations

  • 3 temperatures
  • 5 purification optimization
  • 9 induction conditions
  • 5 extraction methods
1350
Tests

Best expression condition selection

200 media screening
5 best fermentation conditions

1875 Tests

Our high throughput protein production process

StepContentTimelineDeliverables
Host/vector pair determinationGene synthesis including codon optimization
Subcloning in expression vectors
Purification of the best results
To be determined depending on the projectGene constructs
Detailed report including the tests performed
Purified protein from purification test
Protein expression optimizationCulture condition optimizationTest sample if required
Detailed report
Protein purification optimizationDetermination of best purification strategyPurified protein
Detailed report
Process developmentOptimization of bioreactor operating parameters
QC analysis
Options
  • Tag elimination via enzymatic cleavage
  • Endotoxin assay and removal
  • Scale up production / Fermentation

Influence of several parameters on protein expression

Production of difficult-to-express proteins requires varying several parameters that can strongly influence the final yield, purity, stability, solubility and biological activity. If we take a closer look at the productivity more specifically, here are some of the parameters to consider for protein expression optimization:

 

Expression system

The choice of the best expression system is of primary importance when it comes to protein expression. This choice will depend on your requirements: high productivity, proper protein folding, “human-like” post-translational modifications or a compromise between all of these requirements… Here is a small table explaining the essential parameters to consider when choosing the right expression system:

 

Gene sequence

Gene synthesis should be optimized for the host organism by the so-called codon optimization process. This optimization aims at avoiding rare codons in the mRNA sequence thus limiting the risk of protein expression failing and increasing the chance to get a high yield protein production.

 

Vector

Protein expression optimization is also partly driven by the choice of the best expression vector. The non-coding sequence part regulates several elements essential to protein expression:

  • The promoter has to be adapted to the nature of the protein to express. For toxic proteins, promoters with low basal expression will be preferred. Strong promoters are privileged when high yield protein expression are required.

  • The terminator regulates the mRNA turnover and increases mRNA stability in the cytoplasm.

  • The origin of replication controls the plasmid copy number. Choosing a high copy plasmid can be an option if you face low yield protein expression.

 

Expression conditions

Several factors are known to influence protein expression:

  • Temperature: in prokaryotic expression systems such as E. coli, protein expression results in incomplete protein folding and in the formation of inclusion bodies. Even if those inclusion bodies can be exploited, inducing protein expression at lower temperature allows for higher solubility.
    Hypothermic protein expression in mammalian cells results in higher protein expression yields.

  • Concentration of inducer: lowering the concentration of inducer leads to reduced transcription rate thus limiting the formation of aggregates (inclusion bodies) and increasing protein solubility..

  • Media: the growth media should contain all the necessary nutrients to favor recombinant protein expression.

This list is non-exhaustive and represents only part of the parameters ProteoGenix can optimize thanks to its high throughput protein expression service. Contact us to talk about your specific requirements with our PhD account manager who will define with you the best parameters to vary to optimize your protein expression.