Throughput-scalable manufacturing of SARS-CoV-2 mRNA lipid nanoparticle vaccines

Abstract

Lipid nanoparticles (LNPs) are a potent delivery technology that have made it possible for the recent clinical breakthroughs in mRNA therapeutics and vaccines. A key challenge to the broader implementation of mRNA therapeutics and vaccines is the development of technology to produce precisely defined LNP formulations, with throughput that can scale from discovery to commercial manufacturing and meet the stringent manufacturing standards of the pharmaceutical industry. To address these challenges, we have developed a microfluidic chip that incorporates 1×, 10×, or 256× LNP-generating units that achieve scalable production rates of up to 17 L/h of precisely defined LNPs. Using these chips, we demonstrate that LNP physical properties and potency in vivo are unchanged as throughput is scaled. Our chips are fabricated out of silicon and glass substrates, which have excellent solvent compatibility, compatibility with pharmaceutical manufacturing, and can be fully reset and reused. SARS-CoV-2 mRNA-LNP vaccines formulated by our chips triggered potent antibody responses in a preclinical study. These results demonstrate the feasibility of directly translating microfluidic-generated LNPs to the scale necessary for commercial production.

Microfluidic synthesis of lipid-based nanoparticles for drug delivery: recent advances and opportunities

“…232 Further scaling of this platform was recently reported using a 256-element silicon/glass mixer array enabling LNP synthesis at exceptionally high total flow rates up to 283 mL min −1 . 233…”

Nucleic Acid Delivery Nanotechnologies for In Vivo Cell Programming

“…First, as with all encapsulated carriers, LNPs improve cargo stability by preventing encapsulated nucleic acids from deteriorating and being broken down by nucleases, extending their shelf life and improving bioavailability . Second, LNPs are comparatively simpler to manufacture than viral vectors due to advances in scalable microfluidic mixing methods that permit liter-scale development of LNPs. , Third, LNPs are amenable to redosing, which makes them especially helpful for immunization campaigns and transient dosing therapeutic regiments . In terms of challenges and disadvantages of LNPs for gene delivery, conventional LNPs administered intravenously might not successfully target particular tissues or cell types due to limited tropism outside of hepatocytes, which could reduce their efficacy in some therapeutic applications .…”

mRNA Lipid Nanoparticles for Ex Vivo Engineering of Immunosuppressive T Cells for Autoimmunity Therapies

“…Each excipient assists with overall LNP structure and stability; specifically, DOPE strengthens the lamellar structure and promotes endosomal escape, cholesterol enhances LNP membrane stability, and PEG allows for reduced LNP aggregation. ,−, The molar ratio of excipients35%, 16%, 46.5%, and 2.5% of ionizable lipid, DOPE, cholesterol, and PEG, respectivelywas selected based on an optimized LNP formulation previously shown to potently deliver mRNA . We solubilized these four lipid components in ethanol and combined them with mRNA using a microfluidic mixing device with a staggered herringbone mixer structure to formulate our LNP library for screening (Figure A). , An additional LNP formulated with C12-200 ionizable lipidshown previously to be a highly potent ionizable lipidwas used as a positive control, resulting in a complete library of 19 LNPs. , …”

“…67 We solubilized these four lipid components in ethanol and combined them with mRNA using a microfluidic mixing device with a staggered herringbone mixer structure to formulate our LNP library for screening (Figure 2A). 68,69 An additional LNP formulated with C12-200 ionizable lipid� shown previously to be a highly potent ionizable lipid�was used as a positive control, resulting in a complete library of 19 LNPs. 70,71 We characterized the LNP library on the basis of several physicochemical properties that have been shown to impact delivery efficacy.…”