@@ -567,7 +567,7 @@ export function Engineering() {
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<H3text="Interation 4 - Spray-dried SORT LNP called Airbuddy"id="it4"/>
The next design iteration incorporated the insights from Wang's LNP work for building upon SORT principles to make the nanoparticles lung-specific [6]. The main components include DMG-PEG 2000, cholesterol, DOPE and DOTAP as phospholipids and cationic lipids such as 4A3-SC8. In our LNP development, we carefully considered the use of PEG. While PEG can improve stability, it can also reduce cellular uptake and induce immune responses, necessitating a balanced approach to its inclusion [7].
<Collapsibleid="Col1"open={false}title="Ambivalence of PEG and our implementation // LNP Handbook Cooperation">
<Collapsibleid="Col1"open={false}title="Ambivalence of PEG and our implementation">
<p>
<H4text="What is PEG and why is it important for LNPs?"id="text"/>
Polyethylene glycol (PEG) is an essential component in the formulation of lipid nanoparticles (LNPs), which are widely used in drug delivery systems, particularly for mRNA-based therapies like vaccines. PEG-lipids are hybrid molecules consisting of a hydrophilic PEG chain attached to a hydrophobic lipid anchor. This unique structure enables PEG-lipids to interact effectively with both aqueous environments and lipid structures, such as cell membranes and lipid nanoparticles themselves.
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@@ -579,9 +579,7 @@ export function Engineering() {
<p>We collaborated with <aonClick={()=>goToPagesAndOpenTab('corden','/human-practices')}>Corden Pharma</a>, a specialist in LNP technologies, to address these concerns. Based on their recommendations, we opted for <strong>mPEG-2000-DSPE</strong> as our PEG-lipid of choice. This variant minimizes cytotoxicity while providing excellent stability and circulation time. It has also proven effective in reducing immune-related side effects while preserving the integrity and performance of our nanoparticles. </p>
<H4text="DMG-PEG2000 vs mPEG-2000-DSPE"id="text"/>
While mPEG-2000-DSPE has traditionally been used for stabilizing LNPs and enhancing delivery efficiency, we decided to incorporate DMG-PEG2000 into our SORT LNP-based AirBuddy due to its superior properties. DMG-PEG2000 offers better biodegradability and enhanced stability in pulmonary applications. Unlike mPEG-2000-DSPE, which tends to accumulate in the body and may lead to immune activation over time, DMG-PEG2000 is known for its faster clearance and reduced potential for long-term toxicity. For lung-specific delivery, where stability and safety are critical, DMG-PEG2000 ensures that the nanoparticles remain stable long enough to deliver the therapeutic material effectively, but also degrade at a rate that minimizes unwanted immune responses. This makes DMG-PEG2000 a more suitable choice for therapies targeting CFTR-related diseases, where precise and safe delivery to the lungs is essential for treatment success.
<p>Details about the biosafety aspects of our LNP can be read <aonClick={()=>goToPageAndScroll ('sort-lnp-and-cytotoxicity','/safety')}> here </a>. </p>
<p>Details about the biosafety aspects of our LNP can be read <aonClick={()=>goToPageAndScroll ('sort-lnp-and-cytotoxicity','/safety')}> here </a>. </p>
<H4text="Conclusion"id="text"/>
We use DMG-PEG2000 in our SORT LNP-based AirBuddy because of its superior biodegradability, enhanced stability, and reduced risk of immune system activation. By building on insights from experts and incorporating principles from Wang’s LNP work, we’ve tailored our nanoparticles for lung-specific delivery. This choice ensures that our formulations remain stable long enough to deliver the therapeutic payload effectively while minimizing potential long-term toxicity. This balance is crucial for pulmonary applications, where DMG-PEG2000 outperforms alternatives like mPEG-2000-DSPE, making it the ideal choice for treating CFTR-related lung diseases.
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Schematic view of our lung-specific SORT LNP called AirBuddy.
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<Collapsibleid="Col2"open={false}title="Composition of our SORT LNP called Airbuddy">
<Collapsibleid="Col2"open={false}title="Composition of our SORT LNP called Airbuddy // LNP Handbook Cooperation">
<H4text="Components of AirBuddy"id="text"/>
<H5text="Ionizable Lipid"id="text"/>
<p>The primary ingredient, 4A3-SC8 or MC3, are ionizable cationic lipids that forms the core of the LNP. Ionizable cationic lipids become positively charged in acidic environments, such as when a pH change occurs for example in acidic buffers or in the endosome. This allows them to bind to negatively charged nucleic acids and form protective capsules around it. In the endosome these lipids facilitate endosomal escape through electrostatic interactions between the LNPs and the endosomal or cellular membranes.</p>
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@@ -607,7 +605,9 @@ export function Engineering() {
<p>DMG-PEG (Dimyristoylglycerin-polyethyleneglycol) is an important component by improving the LNP stability and preventing aggregation of the LNPs. </p>
<H4text="Production Methods"id="text"/>
<H5text="LNP Assembly"id="text"/>
<p>Our LNP can be formulated using various methods depending on the scale of production, including pipette mixing, vortex mixing, or microfluidic mixing. After mixing the lipids with mRNA in carefully controlled ratios, the mixture is typically dialyzed to remove organic solvents like ethanol and citrate buffer. The choice of lipid composition and preparation method influences the tissue-targeting capabilities of the LNPs, allowing for selective delivery to organs like the liver, lungs, or spleen. For more detailed information on formulation methods and lipid selection, refer to our LNP Handbook deigned in <aonClick={()=>goToPageAndScroll ('handbook','/human-practices')}> cooperation with iGEM Team Linköping </a> and others.</p>
<p>Our LNP can be formulated using various methods depending on the scale of production, including pipette mixing, vortex mixing, or microfluidic mixing. After mixing the lipids with mRNA in carefully controlled ratios, the mixture is typically dialyzed to remove organic solvents like ethanol and citrate buffer. The choice of lipid composition and preparation method influences the tissue-targeting capabilities of the LNPs, allowing for selective delivery to organs like the liver, lungs, or spleen. For more detailed information on formulation methods and lipid selection, refer to our LNP Handbook deigned in <aonClick={()=>goToPageAndScroll ('handbook','/human-practices')}> cooperation with iGEM Team Linkoping </a> and others.</p>
<p>By combining these components with the spray drying method from <aonClick={()=>goToPageAndScroll ('rnhale','/human-practices')}> RNhale </a> [2] we offer a versatile and efficient method for delivering mRNA therapeutics to the lung, paving the way for gene therapy, especially our Prime Guide. The effective delivery of the prime editing complex is a crucial point in our project. </p>
