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

    Step Content Timeline Deliverables
    Host/vector pair determination Gene synthesis including codon optimization
    Subcloning in expression vectors
    Purification of the best results
    To be determined depending on the project Gene constructs
    Detailed report including the tests performed
    Purified protein from purification test
    Protein expression optimization Culture condition optimization Test sample if required
    Detailed report
    Protein purification optimization Determination of best purification strategy Purified protein
    Detailed report
    Process development Optimization 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.