Your go-to option for producing a new protein. Get proteins with no minimum residues in a remarkably simple and scalable way—with no columns, no beads, and no protease.
The PandaPure Protein System streamlines expression, purification, and tag removal, turning traditionally complex procedures into the simplest of operations.
Significance: It's better to get proteins with the exact native sequence you want for research and applications. So why is it often avoided? Because traditional workflows relies on affinity tags are a painful cycle of multi-step chromatography, slow dialysis, and finicky protease treatments to remove them. Now, thanks to PandaPure Protein Systems that shift the heavy lifting to biology, your first choice can always be using proteins as you designed.
Say goodbye to loading, binding, eluting... and all the tedious operations. PandaPure Protein systems combine integrated biological programs with a simple bench protocol.
Clone your protein of interest (POI) into the vector and co-transform it with the organelle plasmid into the host cells.
A receptor on the synthetic organelle and a corresponding signal on the POI enable specific protein translocation.
An inducible cleavage site is placed between the signal and the POI for tag removal.
Grow your cells and induce co-expression.
Synthetic organelles are built de novo via self-assembly and phase separation.
Organelles autonomously bind and sort the POI away from host cell components.
Open the cell envelope, release host components, and isolate the organelles.
The organelles are large and dense, and can therefore be easily separated through simple centrifugation or filtration.
Incubate with the PandaPure™ Protein Reagent to cut the tags, then separate the products in one final spin.
The reagent provides a mild condition to induce the hydrolysis of the cleavage site.
The tag-less protein is then released from the large organelles and recovered through simple separation.
See an example workflow of protein expression, purification and tag removal using our PandaPure Protein System for E. coli.
Co-transform the organelle plasmid (up) and your protein expression plasmid (bottom) into an E. coli host, like BL21(DE3).
Induce expression. The cell produces your target protein and the organelles, which separate it from impurities autonomously.
Harvest and open the cells. Then, in one simple separation (like centrifugation), you collect the target-enriched organelles from the raw lysate.
Use the PandaPure™ Protein Reagent for tag cleavage. Then, with one last spin, recover the pure, tag-free protein out of the organelles.
First, program your cells. Simply co-expressing genes for your designed targets and organelles. See below an example of two-plasmid system offered by PandaPure Protein Expression and Purification Kits‡.
Then perform expression and purification, only requiring several rounds of liquid handling and separation by centrifugation. See below an workflow of performing PandaPure in vivo in E. coli .
To make your life easier. We are dedicated to pushing the boundaries of PandaPure to meet diverse needs. You can access our growing collections via PandaPure Kits – and stay tuned for more upcoming offerings!
PandaPure makes it simpler than ever, at least 3X fewer units operations than a traditional "one-step" method, much more if multi-step affinities are involved.
Anyone can learn PandaPure easily! We've helped computer scientists, environmental engineers, and biology hobbyists successfully learn and use PandaPure to obtain tag-free, purified proteins within weeks.
PandaPure saves your time! The streamlined PandaPure workflow drastically cuts down active bench time, resulting in ~ 85% reduction in hands-on operations compared to typical batch protocols.
PandaPure can act in 96 wells. Parallel processing is easily enabled in standard micro-well plate formats. The liquid handling steps are compatible with automated platforms as simple as an OT-2, enabling efficient screening of large libraries (e.g., mutation variants, expression conditions).
The core PandaPure principles apply to larger volumes. The system is compatible with standard bioreactor processes, simplifying process development and pilot production without extensive protocol re-design.
In contrast, tradition methods are complex and diverse. Unlike PandaPure, a list of very distinct workflows with are needed to handle different applications and scales, while using different consumables from batch resins, high-pressure columns, to magnetic beads.
PandaPure requires no extra investment. Even if you need to purchase a new centrifuge, it still costs a fraction of an entry-level FPLC machine.
Chromatography consumables also cost a lot. In a typical process, affinity columns and external enzymes are responsible for 90% of the cost. PandaPure turns these costs into a one-time investment of the bioprogram.
Save >$300 per mg protein♠. Eliminate the major costs associated with chromatography columns (or magnetic beads), additional proteases, and dialysis consumables, PandaPure can significantly reduce project budgets, even at relatively small scales.
Reduce waste of 10,000. Downstream processing (DSP) contributes significantly to waste in bioproduction. PandaPure's shortened procedure offers substantial environmental benefits. See below the example of comparison for protein purification and tag removal.
For typical protein expression and purification, Process Mass Intensity (PMI) can drop dramatically (e.g., from ~11,000 to ~500), reducing solvent use and waste, especially the need for washing and dialysis.
Inspired by natural organelles, which are capable of sorting, separating, and storing biological molecules within the living cells.
We built synthetic organelles or equivalent, to capture targets of interest with high performance, specificity and efficiency.
Synthetic membrane-less organelles can be built de novo from biological molecules (e.g., proteins, RNAs, and DNAs), and engineered for various properties. The key driving forces for their formation of synthetic organelles include molecular self-assemblies of ordered structures and liquid-liquid phase separation of fluidic systems.
Example: PandaPure Protein System for bacterial E. coli
Preparing PandaPure Organelle plasmid to express synthetic organelle (up), and cloning the target gene into the cognate PandaPure Expression Vector (bottom). Co-transforming two plasmids into a expression host, e.g., E. coli BL21(DE3).
Induce co-expression of organelles and targets with respective inducers (e.g., aTc and IPTG), or via auto-induction medium.
Open the cell structures by disrupting cell membrane and/or cell walls, to isolate organelles enriched with target proteins. If necessary for higher purity, organelles are further washed with proper buffers.
React the organelles with PandaPure™ Protein Reagent, to mediate tag cleavages. Then recover the tag-less proteins in a simple centrifugation and/or filtration.
