<QaBox q="Can you specify the shelf life for spray-dried LNPs? What storage conditions do you recommend to maximize stability?" a="For small interfering RNA, we ofhave demonstrated and published stability of 18 months at room temperature. However, there are no existing studies for mRNA, which tends to be more fragile. For optimal stability, we recommend storing dried LNPs in a cool, dry place, away from direct sunlight."/>
<QaBox q="Can you specify the shelf life for spray-dried LNPs? What storage conditions do you recommend to maximize stability?" a="For small interfering RNA, we have demonstrated and published stability of 18 months at room temperature. However, there are no existing studies for mRNA, which tends to be more fragile. For optimal stability, we recommend storing dried LNPs in a cool, dry place, away from direct sunlight."/>
<QaBox q="What technical requirements and equipment are necessary to successfully spray-dry LNPs using your nano-embedded-microparticle (NEM) technology?" a="The key equipment includes an Impingement Jet Mixer for the preparation of LNPs and a spray dryer for formulating the dry powder. In combination with standard laboratory equipment such as mixers, pipettes, etc., this setup is sufficient to produce NEMs as described in our publications. However, to monitor the process and assess the quality of both intermediate and final products, a variety of analytical methods is required, including Dynamic Light Scattering (DLS), Nanoparticle Tracking Analysis (NTA), Plate Reader, cell culture equipment, and a laser diffractometer. This setup makes it quite costly to establish the entire process."/>
<QaBox q="How complicated is the protocol for producing spray-dried LNPs? Can you describe the protocol?" a="The spray-drying process for LNPs using our NEM technology involves several technical steps. Initially, a mixture of an ethanol phase containing lipids and another phase with RNA is prepared. This mixture is then subjected to an Impingement Jet Mixer to formulate RNA-loaded LNPs. Subsequently the LNPs are mixed with a stabilizing excipient. This mixture is fed into a spray-drier where the solution is aerosolized into a heated drying tower. The liquid evaporates, leaving behind NEMs as small spherical particles, which can then be loaded into capsules for use with a dry powder inhaler (DPI)."/>
<QaBox q="How do LNPs appear after the drying process?" a="In the dry state, LNPs cannot be visualized, as they are embedded within the NEM structure. However, we assess the LNP structure and morphology after redispersion of the NEMs, a process that also occurs upon inhalation and contact with lung fluids. While we observe a slight increase in LNP size after redispersion, the particles remain intact, retaining their spherical shape and RNA loading, as demonstrated by various techniques, including cryo-TEM imaging."/>
<QaBox q="Does each LNP formulation require individual testing?" a="Yes, each LNP with a unique composition needs to be tested individually to ensure optimal stability and performance."/>
<QaBox q="How can we test the efficiency of LNPs after the drying process?" a="The efficiency can be tested through transfection studies. Additionally, the particles can be broken down to analyze the mRNA structure, although this is a more complex and time-consuming process."/>
<QaBox q="Can you elaborate on how you use AI to customize LNPs? What exactly does the AI do, and how reliable is it?" a="Our AI is used for screening, optimization, and the design of experiments, significantly reducing wet lab work. It also plays a role in developing new lipids, a process more closely associated with the work of Olivia Merkel."/>
<QaBox q="In your opinion, which LNPs are best suited for the drying process (SLNs, NLCs, etc.)?" a="It depends on the specific application and composition of the LNPs."/>
<QaBox q="What properties of LNPs could hinder the drying process?" a="Theoretically, nothing should hinder the drying process if it is optimized for the specific cargo and target. Adjustments can always be made to accommodate different formulations."/>
<QaBox q="What are the estimated costs for the entire drying process?" a="The primary expenses are in raw materials and for the formulationspecifically RNA and lipids, which are relatively expensive. Other consumables, such as process fluids, are comparatively inexpensive. The manufacturing process itself is cost-effective; however, overall expenses will vary based on production volume and equipment utilization rates."/>
<QaBox q="Would you be willing to support us in our project? Would you dry our LNPs?" a="The spray dryer requires 5 mL of a solution with 5% solid content. Among these solids, the lipid content can be adjusted between approximately 0.2% and 15%, while the RNA content can range from 0.02% to 1.5%. The remaining percentage is composed of excipients. We’ve published recovery rates of >70%. You can send us the mRNA and LNP components to encapsulate and dry."/>
