HIGH THROUGHPUT PROTEIN EXPRESSION TESTS

Our high throughput protein expression test service 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.

High throughput protein expression test
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:

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

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.