How are lyophilised beads manufactured?

Lyophilised reagent beads are durable spheres of freeze-dried material formed from accurately measured and dispensed volumes of customisable formulation.

These single-dose beads – like all lyophilised material – retain long-term stability at ambient temperature and therefore do not need expensive cold chain shipping or refrigeration during storage.

This significantly increases the accessibility of diagnostic testing for difficult-to-reach patients, facilitating regular, near-patient testing in the community.

Lyophilised beads also present further performance and efficiency opportunities, particularly in terms of assay accuracy and speed and ease of process.

  • Accuracy – Lyophilised beads contain a single, precisely measured dose, minimising the potential for variance.
  • Cost savings through improved efficiency – With less risk of user errors, a longer shelf life, and less exposure to moisture, wastage is significantly reduced, saving time and money.
  • Quick and simple – Lyo beads reconstitute quickly due to their volume to surface area ratio and come pre-packaged, ready for immediate use.

The process of manufacturing lyophilised (lyo) beads requires a high level of expertise, experience and equipment. Partner with a contract manufacturer like Biofortuna and you can deliver the benefits of lyophilised beads without prohibitive start-up costs, wasteful trial-and-error testing, or risk to your capital.

Looking For Expert Technical Advice? Contact Us.

Biofortuna has leveraged years of experience to create a sophisticated lyo bead manufacturing process, that is designed to produce a perfectly accurately measured dose in every bead.  Here’s how it works…

Formulation optimisation

To create a custom lyophilised bead which is stable and suitable for large scale manufacture, the ideal freeze-drying conditions of the assay must be established before the lyophilisation process begins. This is usually achieved by the combination of Freeze-Drying Microscopy (FDM) and Differential Scanning Calorimetry (DSC).

FDM uses a miniscule amount of a complete formulation to freeze the sample and put the chamber under vacuum. These conditions are accurately controlled, reproducing those of a manufacturing freeze drier. Observing the crystallisation, freezing and collapse temperature of the sample can help identify optimal temperatures, challenging reagents and the effect varying concentrations of the excipients can have on the final conditions.

DSC can calculate how the Glass Transition Temperature (Tg’) will be affected under different heating rates of the product. This provides an indication of the duration of the lyophilisation cycle. This helps to calculate the benefit that annealing (Thermal Treatment) of the sample could have on the final conditions.

Both FDM and DSC will be performed with a range of varying formulations identifying challenging components and the most promising excipient combinations in varying concentrations. This provides a good understanding of the conditions to be applied on a larger scale with a minimum reagent cost to the client.

Scientists putting assays into long freeze dryers

Scientists putting assays into long freeze dryers

Automated dispensing

Once the ideal freeze-drying conditions of the assay are established, accurately measured droplets of this formulation are dispensed into liquid nitrogen. This accuracy will ensure that each lyo bead contains a precision dose for its final application.

The beads are transferred to lyophilisation freezers to begin the next stage.

Lyophilisation

Lyophilisation takes place in two cycles – primary and secondary drying.

Primary drying cycle (sublimation)

During sublimation, the pressure in the environment is lowered while a vacuum is applied. This causes the ice to sublime, removing 95% of the water.

Secondary drying cycle (desorption)

Desorption will remove the remaining, unfrozen water molecules from the formulation. An inert gas is then used to break the vacuum in the environment, ready for the product to be sealed.

Quality control

Biofortuna will always ensure the highest level of quality in each product before the process is complete. This happens in a controlled humidity environment, where the formulation is tested to meet the required standards for appearance, size and successful drying.

First, the Karl Fischer titration technique is applied to detect the levels of residual moisture.

Successful freeze-drying will have removed most of the water from the formulation and the Karl Fischer test allows us to measure the percentage of moisture left in the dried product. However, more is not always better. There is a fine balance between a product that has been optimally dried and one that has been overdried. Depending on the formulation, excessive dryness could have a negative impact on the assay efficiency and/or the bead’s physical characteristics. It takes a certain expertise to fine-tune this balance.

The beads are then evaluated on their physical properties. This includes ultimate tensile strength (maximum strength a bead can withstand before breaking) and friability (how easily the beads can break into smaller parts under duress).

These are important properties that need to be well defined, especially if there is further manipulation after the manufacturing of the beads, such as bulk transport, or placement into cartridges/microfluidic devices.

Beads are also checked for their dissolution rate. This is a critical property, especially when the beads are to be used in a POC (Point of Care) device, where timing from the sample/buffer injection to the final result are very stringent.

Finally, beads are scored on visual appearance, such as their texture, roundness and consistency. This is usually done for aesthetic reasons, but the presence of irregularities could provide indications on the robustness of the freeze-drying process.

The final part of the Quality Control process is, of course, to test the final product for assay sensitivity. This can be done either at the bead manufacturer’s site or the customer’s site. When the sensitivity of the assay is proven to be within the detection limits defined, then the product can be released.

Why Biofortuna?

We partner with some of the world’s most innovative assay developers, supporting them to take their assays from concept through to commercialisation.

Our support is tailored around your exact requirements and defined by a problem-solving mentality. We listen to precisely what you need and find the solution to match.

A combination of market-leading expertise, ISO 13485 and ISO 17025 accreditations, FDA registration and state-of-the-art facilities mean that you can trust us to employ the highest scientific standards when bringing your product to market.
See Biofortuna's manufacturing process in this infographic
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