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Design your molecular assay using lyo-ready or custom lyophilized Applied Biosystems™ TaqMan® Master Mixes
Strategic partnerships can be critical to the success of great ideas
The unfolding of the COVID-19 pandemic is a great example of how the benefits of strategic partnering can yield unprecedented innovation. Quick leveraging of resources across the globe enabled the discovery of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as the causative reagent of COVID-19 leading to SARS-CoV-2 testing and detection in millions of people.
A distinct outcome from SARS-CoV-2 testing and detection is the affirmation that quantitative reverse-transcription polymerase chain reaction (RT-qPCR) is a powerful tool.
An additional conclusion from global SARS-CoV-2 research is a reinforced understanding that easily available molecular tests are critical to address geographical and accessibility constraints of the world’s patient populations.
Research laboratories have used both RT-qPCR and quantitative PCR (qPCR) assays for decades to detect the presence of nucleic acids and gene expression changes in human, animal, and plant biological samples. Using fluorescent reporter probes, DNA can be quantitated in real time using qPCR. Adding a reverse transcriptase to transcribe RNA into complimentary DNA (cDNA), RT-qPCR can measure specific levels of gene transcripts.
New molecular tests and applications have sustained the demand for lyophilized qPCR or RT-qPCR reagents even as the molecular testing industry sees declines in testing for SARS-CoV-2.
The good news if you are developing molecular assays is that numerous options are available to secure custom & catalogue raw material manufacture (CRM) lyo-ready or lyophilized qPCR and RT-qPCR reagents. Some molecular test developers may even choose to outsource the entire manufacture of lyophilized reagents to help accelerate bringing their molecular assays to market.
Custom lyophilized Applied Biosystems™ TaqMan® assay technology
Thermo Fisher Scientific has long been a trusted provider of TaqMan® assay technology for researchers needing qPCR and RT-qPCR solutions.
TaqMan® qPCR and RT-qPCR master mixes include fast-activating DNA polymerases and thermostable reverse transcriptases that are compatible for fast cycling, high sensitivity, and multiplexing capabilities all combined with reliable performance.
What is less known is how the Thermo Fisher Genetic Sciences OEM and Commercial Supply team can partner with businesses of all sizes and in all stages who could benefit from incorporating a custom lyophilized solution into their molecular assay development process.
Here are some considerations for lyophilized assay development and three options for how Thermo Fisher OEM can partner to help molecular assay developers power their test design with custom lyophilization.
Why lyo-ready or lyophilized master mixes?
As mentioned earlier, a molecular assay increases its impact when it can be used off the shelf when and where a test is needed. The convenience and stability of ambient tests result directly from the benefit of reagents that are dried-down or lyophilized eliminating the need for more expensive cold storage.
Molecular tests that include lyophilized reagents also help reduce mixing and handling steps to increase the total number of tests that can be run at one time.
As opposed to wet assay reagents, which may need to be separately dispensed and frozen, lyophilization offers the possibility of combining assay components into one convenient test mixture.
Key considerations for lyophilizing reagents
While lyophilization provides great benefits at the point of use, offering a ready-to-use molecular test means a bigger burden rests on the test provider to manufacture a quality product.
The lyophilization process, which uses freezing and vacuum to dehydrate biological samples through sublimation, involves many factors to ensure a dried-down reagent or biologic performs in the final assay as intended.
- One major consideration is scale. The equipment needed to lyophilize can range in cost from thousands to millions in investment. Freeze dryers, vacuum pumps, and sample holders all must meet the requirements of the sample type, format, and scale of the samples that need lyophilization.
- Another consideration is the sample formulation. Buffers for reagent components that work in a standard wet assay may not be compatible for the lyophilization process. For instance, 10% to 50% glycerol is a common cryoprotectant added to stabilize PCR enzymes stored frozen at -20°C. Similar glycerol percentages included in an enzyme formulation for lyophilization would result in a sticky dried-down clump.
- A big pain point when lyophilizing is determining a proper formulation of active components and excipients to ensure an assay performs similarly pre and post lyophilization. Excipients are inert components added to assay formulations that serve specific purposes such as binding, preserving, bulking, emulsifying, etc. all to support the assay effectiveness.
- A final consideration is the quality control (QC) throughout the lyophilization process. Molecular assays adhere to more rigorous compliance standards than basic research assays. Lot-to-lot variations in assay formulations need to be verified and manufacturing processes validated as part of numerous standard certifications like ISO 13485.
The Thermo Fisher OEM team can help with all these key considerations depending on the molecular assay developer’s needs.
Let’s look at three OEMpowered options for integrating CRM lyo-ready or custom lyophilized TaqMan® qPCR and TaqMan® RT-qPCR master mixes.
Custom OEMpowered Option 1
Experienced molecular assay developer
Molecular assay developers who already have established lyophilization processes for their assays may want to keep their manufacturing process proprietary or manage cost per assay. In this case, it doesn’t make sense to bring in an OEM partner to help develop the lyophilization procedure or quality control process.
This established test developer still may need a more reliable source for the qPCR or RT-qPCR components of their test. Variations in the supply chain are always possible or the quality of their current enzyme formulations may be frequently failing required QC specifications.
For established assays a great option would be to source new TaqMan® Lyo-ready reagents to replace qPCR or RT-qPCR assay components that are not meeting product requirements.
See product offerings (Table 1, Option 1) for assay developers looking to switch to a more reliable Lyo-ready TaqMan® qPCR or RT-qPCR master mix.
Custom OEMpowered Option 2
Molecular assay developers new to lyophilization
Some molecular assay developers have a great wet assay but have limited lyophilization experience.
These assay developers may want to first establish a new lyophilization process on their own or want to lyophilize their assay at a smaller scale before committing to a large-scale formulation or equipment costs.
Molecular assay developers with limited lyophilization experience could benefit from initial lyophilization guidelines and TaqMan® qPCR or RT-qPCR formulations that already include pre-verified excipients.
See product offerings (Table 1, Option 2) for assay developers looking for more lyophilization assistance.
Custom OEMpowered Option 3
Molecular assay developers with a new idea looking for a lyophilized solution provider
For molecular assay developers seeking a turnkey assay development solution, a third option is to outsource the entire customization and manufacture of the assay to one provider.
The benefit of partnering with a reagent provider for a complete custom lyophilized solution is that the onus of quality control and manufacturing rests on the contract manufacturing organization (CDMO). A good CDMO will be able to offer a variety of assay formats and to ensure an assay is produced according to ISO 13485 standards.
While this route may incur more cost upfront and has starting volume requirements, working with a trusted OEM partner like Thermo Fisher means the final custom lyophilized assay will have high sensitivity, reproducibility, and lot-to-lot consistency.
See product offerings (Table 1, Option 3) for assay developers looking for a turnkey lyophilization solution.
Table 1. OEMpowered custom lyophilization options*
Even if the three options discussed here don’t quite fit your needs, the Thermo Fisher OEM and Commercial Supply team has the flexibility to address any custom lyophilization request.
Thermo Fisher also can assist clients with repackaging or reconfiguring a technology, licensing intellectual property to bring a technology to market in a unique way, or solving global market challenges.
Themo Fisher understands that the right OEM partner adds value, and our goal is to bring outstanding value to our customers.
Connect with a business development manager to learn more
Related Links & Resources
- Learn more: Genetic Sciences OEM and Commercial Supply
- Article: Four Things You Should Know About Customized OEM Solutions
- Article: How to Help Ensure Your Supply Chain Security
- Article: Nine Questions to Ask When Considering an OEM Partner
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