Our mission is the stabilization of sensitive biochemical and molecular reagents for applications in diagnostics and research. The core technology for this purpose is freeze-drying, also referred to as lyophilization – a field where we at Appylo have extensive experience in development and production. But we also offer other methods of reagent stabilization.

• About Lyophilisation (Freeze-Drying)
• Freeze-Drying in the pharmaceutical industry
• Freeze-Drying in diagnostic applications
• Freeze Drying of reagents
• Freeze-Dried Applyo Beads
• Applyo’s Freeze Drying technology
• Freeze-Drying in comparison to other methods for solid and stabilized reagents

About Lyophilisation (Freeze-Drying):

Freeze-drying is a dehydration process through sublimation, i.e. the phase transition of water from solid directly to vapour. This is possible when temperature and pressure are below the triple-point in the phase diagram of water. Below that point, water is gently removed from a frozen aqueous sample by sublimation. Simultaneously, the molecules are stabilized by lyoprotectans, polymers and other excipients which conserve the shape of the product and facilitate fast resuspension. Therefore lyophilization is the method of choice for stabilization of products which contain thermo-instable and expensive substances. Rapid rehydration before application as it is required by mostmolecular biology assays. is assured by the high porous structure of the lyophilizate.

Although the lyophilization process is time and energy consuming, the total CO₂ footprint of freeze-dried : Cold chains for storage and distribution are not required any more.

Freeze-Drying in the pharmaceutical industry

Freeze-drying has become a fast-growing segment in the field of IVD reagents even before the SARS-CoV-2 pandemic. The increasing demand for high-performance reagents makes lyophilization the key technology for supply of high-performance therapeutic drugs, vaccines as wells as enzymes, proteins and nucleic acids. Lyophilization also enables new possibilities for complex reagent formulations that are unstable in liquid form.

Freeze-drying technology typically used in pharmaceutical or diagnostics industry requires the conventional flasks closed by stoppers or plastic reaction tubes which limits upscaling possibilities. We have overcome this common obstacle of freeze-drying technology with our proprietary production technology.

Freeze-Drying in diagnostic applications

With SARS-CoV-2 a transformation in diagnostics from centralized and professional testing to decentral Point-of-Care (PoC) or individual Point-of-Need (PoN) testing has accelerated. For testing outside laboratories and thus in uncontrolled environments by semi-skilled or even untrained users lyophilization is the means of choice: Pre-packed single-portioned, ready-to-use reactions and no need for cold-chains are prerequisites for PoC- or PoN-solutions.

The challenge remains in the design of devices such as cartridge consumables and disposables for easy-to-use kits.

Freeze-Drying of reagents

For freeze-drying of reagents excipients are used to stabilize the products shelf-life and give it  a mechanically stable shape as well as a porous structure for e.g. pick-and-placing and fast resuspension behavior. After the lyophilization process lyo cakes remain in the containers chosen for freeze-drying. These can now be stored and handled at ambient temperature.

Applyo’s proprietary technology renders containers unnecessary for freeze-drying, thereby increasing the freeze-drying capacity of the machines substantially. Furthermore, Applyo Beads are single-apportionable and can be picked and placed as required due to their high mechanical stability.

Conventional freeze-drying approach

• inhomogeneously sized lyo-cakes or beads due to dispensing into liquid litrogen, which boils up
• less control of lyophilization parameters due to processing in plastic material
• expensive due to the use of microtiter plate consumables
• Batch size directly correlated to shelf area in freeze dryers
• Risk of contaminaton by
  • low temperature stress-induced micro-cracks of PCR reaction tubes
  • liquid nitrogen

Applyo‘s freeze-drying approach

• Homogenous spheres in terms of size, volume and assay performance
• Low cost due to no plastic consumables
• Full control of freeze drying parameters possible
• Mechanically stable – optimized for direct transfer with vacuum tweezer
• High range of bead size and volume: from1.6 mm to > 8 mm diameter (2 µl to > 250 µl) as well as  3 ml (~ 18 mm diameter) demonstrated
• Largest bulk-lyophilization batch in market

Other Methods for the production of solid stabilized reagents

We offer also a variety of technologies for air-, heat- and vacuum-drying of reagents as well as powder compaction (tableting) of dry reagents.