Our Technology

Phase separation is an ancient phenomenon evolved by nature to perform highly specific separations in any complex environment.

We develop reagents that combine engineered biopolymers with tunable phase separation with affinity ligands that confer the unique material properties to the biologic target of interest. These reagents are then paired with high performing filtration processes that can improve productivity and quality. The linear scalability of these processes vertically integrate a biologic’s development as its batch size increases 1000-fold from the bench to the clinic to a commercial enterprise.

Our Reagent

The IsoTag™ reagent technology comprises simple protein reagents and straightforward processes that can plug into existing purification equipment along with validated, off-the-shelf consumables. The protein reagents typically comprise two protein modules - a phase-separation module and a binding module - that are precisely engineered for the target biologic of interest. IsoTag™ has been demonstrated to be target agnostic, whether it be a viral vector, antibody, nucleic acid, enzyme, or any other novel biologic, the principles of the platform are the same.

Binding Module: Designed with exquisite specificity for the Target Biologic to rapidly bind in-solution

Robust, easy-to-express, stable ligands
Libraries of scaffolds with >10E9 diversity enable rapid ligand identification for for new applications
In vitro screening procedures provide high throughput, real-world binding data that can quickly identify novel binding properties, including gentler elution conditions, improved target specificity (e.g. full virus particles), and NaOH tolerance
Can be multimerized as needed to achieve capacity, affinity, and avidity requirements

Phase Separation Module: pulls captured Biologic into large pure droplets on command

Modules are custom designed for each intended use case to provide precise performance
Transduce a variety of bulk user inputs such as temperature, salt concentration, and pH into specific separation at the nanoscale
Multiple families of orthogonal modules enable new bioprocessing synergies to push the technology to be even faster and more efficient (e.g. parallel contaminant removal into the opposing phase as the target)

Phase separation capture is a radical departure from existing technology.

Therapeutics are getting larger, more complex, and more effective. Affinity chromatography, in contrast, has become dramatically less efficient

Innovation in downstream development of therapeutics has not kept pace to provide solutions tailored to the unique needs of advanced medicines. With more open pore sizes, membrane and monolith formats have achieved moderate improvements. Isolere’s small protein capture is a radical departure from existing technology.

Try our Tech

(Therapeutic) size Doesn’t matter!

Consistent Binding Capacity no matter the Therapeutics' size!

The foundational principle for chromatographic separation is using a combination of diffusive and convective forces to separate particles based on their interaction with the surfaces of the resin bead. However, this also ensures that as target molecules get larger, capacity will drop off non-linearly, meaning that a particle 10X as large will have >100-fold lower binding capacity on the same resin.

IsoTag™ reagent is a stark deviation from this approach. By relying on the diffusion and convection of a relatively small protein reagent instead of the target, we observe consistent capacity and performance regardless of application.

Soluble State

Well hydrated and sterically hindering in their soluble state, IsoTag™ reagents create a protective shield around the target biologic during the purification process, preventing further contamination and degradation.

Phase Separated State

Creating protective droplets in their phase separate state, IsoTag™ reagents specifically sequester their targets away from contaminants and enzymes. The protein dense environment prevents aggregation and other mechanisms of activity loss.

Simple Execution

Use your existing purification equipment and similar processing techniques but achieve a new level of purity and efficiency.

Self-Scaffolding

Free your process from separate unit operations that purify your target EITHER on size differences OR interaction differences. IsoTag™ simultaneously leverages on command manipulation of size AND affinity.

Recombinant

Our reagents are recombinant proteins that are simple and straightforward to produce in E. coli.

Fully STERILE & Can be manufactured under full cGMP. NO resin/bead; NO chemical conjugation. NO column packing or packing failures!

FAQs

IsoTag™ is a single chain protein reagent that is delivered in a concentrated stock solution, similar to other protein reagents.
Affinity liquid phase separation (ALPS) is the term for the end-user downstream process. There are two variants of the process:
• Affinity liquid phase separation via tangential flow filtration (ALPS-TFF)
• Affinity liquid phase separation via centrifugation (ALPS-CF)
Yes. The assay is validated (by Cygnus Technologies) and we are currently negotiating a supply agreement for commercial production of the kit.
Residuals of IsoTag™ are <200 ng/E13 vgs in the elution buffer, as determined by beta kits.
IsoTag™ transition can be initiated with as little as a 0.1M change in salt concentration, but we typically recommend a 0.5M change to ensure a consistent and robust droplet formation.
For AAV - approximately 2mL - 3mL of formulated IsoTag™ is required per liter of harvest.
For LV - approximately 5mL - 6mL of formulated IsoTag™ is required per liter of harvest.
In our benchmarking studies, we have typically seen similar or improved (>20%) recoveries from the IsoTag™ ALPS process compared to an industry standard chromatography process.
The ALPS-TFF process is highly effective at washing and concentrating product because of the large pore size of the filter (typically 0.2 um and 0.65 um). To enact a consistent filtration process with these open filters we recommend running at a constant flux using permeate flow control mode.
We expect zero toxicity of any IsoTag™ product. We have not seen any cytotoxicity of IsoTag™ in cell culture and we have injected small animal models with viral preps that contain residual amounts of IsoTag™ to no detriment. Analogous reagents have been used in human clinical trials at doses far exceeding residual quantities.
The IsoTag™ ALPS-CF and ALPS-TFF process is highly scalable because of the mechanism of the droplet formation - the droplets form the same size regardless of volume so whether you are trying to process 100 mL or 1000L, the droplet formation and concentration dynamics are the same.
In our development history of adenovirus, adeno-associated viruses and lentivirus, we have seen zero evidence of reduced viral potency that can be attributed to our purification process.
Low speed centrifugation and tangential flow filtration processes.
Our volume-based process is compatible with serum and serum free feed streams.
Specifically, IsoTag™ AAV reagent is compatible with multiple serotypes (1,2,6,8,9,rh10 and PHP.B) and IsoTag™ LV reagent is compatible with any VSV-G pseudotyped vector.