Improved heterologous protein expression platform for production of recombinant proteins in E. coli

The heterologous expression platform has been developed to alleviate some of the difficulty involved with known processes. This robust heterologous expression system will enable the production of proteins with valuable bioactivities. The system can improve on the production of currently sort after proteins as well as enable the production of novel proteins with unknown production challenges. This function can therefore deliver a local supply of proteins used as consumables and help introduce new proteins and products to the market.

The process includes a self-preparation function (self-preparing technology) which occurs without inhibiting cell growth or the expression of the target protein. Cell lysis can be achieved within five minutes and no additional lysis chemicals or specialized equipment.  With this system protein extraction is faster (>85% lysis after 15 min) and 3 times more total protein can be extracted from cells.

Additionally, the process incorporates fluorescent fusion of target proteins for real time monitoring during production. This increases protein solubility and stability while increasing protein yields. Being able to monitor real time production of target proteins allows for quick problem identification and rapid system optimization. Furthermore, being able to visualize proteins provides unprecedented confidence during purification.

Academic and research institutions

Animal product replacement industry

Diagnostic facilities

Pharmaceutical and biopharmaceutical manufacturing

After heterologous protein expression, the E. coli cells harboring the self-preparing tech prepare themselves automatically for the next step in protein purification which is protein extraction. The self-preparing biotechnology simplifies the protein purification process, decreases purification times, reduces costs and boosts yields by improving the extraction efficiency above current standards.

Main advantages

• Simpler lysis protocol that can be performed in minutes (compared to hours) without the need of additional chemicals/enzymes or specialized equipment
• Better extraction efficiency compared to standard methods such as lysozyme
• Faster processing with >85% lysis after one freeze thaw cycle
• Cheaper purifications by excluding the need for lysozyme or detergents
• No detrimental effects on growth of the E. coli host or recombinant protein expression levels

The use of fluorescent fusion provides real time feedback of recombinant protein expression. This allows for identification of problems during production when they occur potentially saving significant costs on unnecessary downstream processing. Furthermore, real time monitoring allows for rapid optimization of production giving the best possible yields. Purification is also aided by being able to visualize target proteins during purification with the potential to speed up the purification process.

Dr Anton Du Preez van Staden

Ross Rayne Vermeulen

Prof Leon Dicks

Prof Carine Smith

Dominic Nicholas

The developed heterologous expression platform greatly simplifies the production process of recombinant proteins expressed in E. coli. This allows for production of recombinant proteins with over 13% reductions in cost while being more than 10% faster (complete process, protein expression from cells is >85% faster). With these cost and time savings valuable proteins used in industry and research can be produced cheaper, better, and faster.

Being able to identify production problems early through real time monitoring can also significantly reduce costs while increasing productivity through rapid optimization.