Developing A Scalable RNA-LNP Drug Product For Clinical Translation

First identified in 2019, the severe acute respiratory syndrome coronavirus (SARS-CoV-2) is an infectious disease that has resulted in a global pandemic with an unprecedented human impact. As a result, a number of messenger RNA (mRNA)-lipid nanoparticle (LNP) vaccines have been developed.

A common need for these RNA-LNP therapies is a highly compliant and scalable manufacturing process. The current state-of-the-art for large-scale production of RNA-LNPs can be divided into 4 key steps: 1) production of RNA drug substance using in vitro transcription (IVT) from a DNA template, 2) formation of the RNA-LNP drug product by mixing an aqueous stream of RNA with lipids in ethanol and subsequent in-line dilution, 3) purification of the bulk drug product to remove ethanol using tangential flow filtration (TFF) and 4) sterile filtration and final fill-finish in vials.

The LNP encapsulation step requires controlled mixing of RNA and lipids and is among the most difficult unit operations to scale up to high throughput rates and large batch sizes. The mixing process is intrinsic to the drug product and impacts the physical properties, potency and toxicity of the drug. Furthermore, it is cost-prohibitive and time consuming to do process development for RNA-LNPs at full scale.

New instrumentation is allowing for highly reproducible and scalable production of RNA-LNPs over a wide range of flow rates and volumes, overcoming key limitations of bulk methods (such as turbulent in-line mixing). In this work, we demonstrate the capabilities and workflow of the Ignite+ using two representative lipid compositions. We provide examples to show how automated in-line dilution can be incorporated within the Ignite+ workflow to replace manual bulk dilution, and how to make the transition from small-scale dialysis or spin filters to tangential flow filtration (TFF) for post-formulation purification.