@@ -534,12 +534,33 @@ export function Engineering() {
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<Collapsibleid="Col2"open={false}title="Composition of our SORT LNP called Airbuddy">
<p>Our lung-specific formulation uses <strong>Selective Organ-Targeting (SORT) LNPs</strong> [1], optimized for stability, targeting, and delivery efficiency. Combined with the <strong>spray-drying technique</strong>, our LNP <strong>AirBuddy</strong> offers an efficient way to deliver mRNA therapeutics to the lungs. The successful delivery of the prime editing complex via inhalation is key to our project.</p>
<H4text="Components of AirBuddy"id="text"/>
<p>erster absatz</p>
<p>zweiter absatz</p>
<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>
<H5text="Helper Lipids"id="text"/>
<p>DOTAP (Dioleoyltrimethyl-ammonium propane) is a cationic lipid that makes up 50 % of the total molar lipid ratio. It plays a crucial role in binding to the negatively charged surface of lung epithelial cells. This enhances transfection efficiency and helps make the LNP formulation more lung-specific, improving targeted delivery. The neutral helper lipid DOPE (Dioleoylphosphatidylethanolamine) enhances endosomal escape by fusing with the endosomal membrane and improves transfection efficiency.</p>
<H5text="Sterol"id="text"/>
<p>Cholesterol, is an important cationic lipid, providing structural stability, fluidity and permeability to the LNPs, thereby improving their overall transfection efficiency. </p>
<H5text="PEGylated Lipids"id="text"/>
<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 cooperation with iGEM Team Linköping and others.</p>
<H5text="Spray drying Procedure"id="text"/>
<p>By combining these components with the spray drying method from RNhale [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>
<H5text="Storage"id="text"/>
<p>The final LNP solution can be stored at 4 °C for a few days. It is recommended to use the formulated LNPs as soon as possible to maintain consistent results. Storage at RT is not recommended. Storage at freezing temperatures is also not recommended unless optimized cryoprotectants are used.</p>
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<p>The final innovation for our LNP to become <strong>AirBuddy</strong> came through consultation with Benjamin Winkeljann from <aonClick={()=>goToPagesAndOpenTab('rnhale','/human-practices')}> RNhale</a>, where the use of spray-drying techniques was discussed. Spray-drying the LNPs, instead of using traditional methods, helped improve stability and eco-friendliness of the product [8]. The spray-dried SORT LNPs demonstrated lower cytotoxicity, and the technique proved effective in maintaining particle integrity.</p>
<p>The final innovation for our LNP to become <strong>AirBuddy</strong> came through consultation with Benjamin Winkeljann from <aonClick={()=>goToPagesAndOpenTab('rnhale','/human-practices')}> RNhale</a>, where the use of spray-drying techniques was discussed. Spray-drying the LNPs, instead of using traditional methods, helped improve stability and eco-friendliness of the product [8]. The spray-dried SORT LNPs demonstrated lower cytotoxicity, and the technique proved effective in maintaining particle integrity. In conclusion, we created a stable LNP for efficient delivery of mRNA therapeutics to the lungs since the successful delivery of the prime editing complex via inhalation is key to our project. </p>
