Xbrane Rhamex System
Production of host-toxic proteins is a problem for many research scientists and biologics manufacturing technologists. In the past, the choice of production system has been limited and most used systems have been struggling with either too low production levels in the cell or production at ‘full-blast’ resulting in protein aggregates, protein misfolding, and host toxicity.
Xbrane Rhamex System -
A rhamnose inducable protein production technology
Xbrane Rhamex System is tightly regulated at the individual cell level but at the same time capable of producing high protein yields. It facilitates production of more bioactive and correctly folded proteins per production run.
Some of the main advantages of Xbrane Rhamex System:
- Ability to express larger amounts of host-toxic and properly folded (active) proteins than other systems.
- E. coli strain independent.
- Unique mechanism of induction allows for a slower, more uniform production of proteins compared to the all-or-nothing induction mechanisms characteristic of other systems.
- Expression levels can equal those of the T7 system but the inducer costs are reduced, providing great value to the end user.
Tight regulation gives users full control over timing the onset of expression.
The technology behind Xbrane Rhamex System
The Xbrane Rhamex System (pRHA) offers very tight control over protein expression. The rhaR and rhaS regulatory genes as well as the rhaBAD promoter are cloned into a variety of different copy number plasmids while replacing the rhaBAD DNA sequence with a multiple cloning site such that any gene of interest can be placed under the control of the rhamnose inducible promoter.
Figure: Schematic view of Xbrane Rhamex System.
Source: BioTechniques (2006)
As described in a publication on the system in BioTechniques, the Xbrane Rhamex System (pRHA) vastly outperformed both the pET (T7 polymerase) system and the arabinose-inducible pBAD system when tested for the ability to produce a functional host-toxic protein in E. coli. The Xbrane Rhamex System’s unique ‘rheostat’ mechanism of induction allows researchers and biologics manufacturers to regulate the production of protein produced by a culture on a per cell level, facilitating the expression of normally toxic proteins or generating greater yields of correctly folded proteins. The amount of protein produced per cell in a culture is directly proportional to the amount of rhamnose added to the culture medium. This is in direct contrast to both the T7 and pBAD systems in where the number of cells expressing at ‘full blast’ in the culture are proportional to the amount of inducer added to the culture medium. Xbrane Rhamex System is very tightly regulated by the addition of glucose to the culture medium, allowing the end user to carefully time the onset of expression.
Link to the publication from BioTechniques>
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