Microfluidics production of self-amplifying RNA-based therapeutics vaccine

Summary: THERAPEUTIC-SAM@SCALE will support the UK Manufacturing strategy to build on the technology developed for COVID-19 self-amplifying RNA vaccines. We will identify the key material and manufacturing variables that must be considered for self-amplifying RNA therapeutic vaccines for cancer. This will be transferred to GMP manufacturing scale and validated in vivo. Through this process we will support the national and international supply of novel thermostable vaccine systems.

Background: In response to COVID-19, Perrie has been working with the UK Government Vaccine Task Force to support the manufacture of a COVID-19 self-amplifying RNA vaccine. RNA-based vaccines combine the positive attributes of both live-attenuated and subunit vaccines. However, these RNA vaccines are only effective when formulated within a nanoparticle delivery system, and lipid nanoparticles (LNPs) has been adopted for the current COVID-19 vaccines. Lipid nanoparticles protect the RNA against degradation, facilitate endosomal escape and cell specific targeting, and can be co-delivered with adjuvants. Furthermore, they have the potential to be manufactured in a scale-independent manner. Given the positive responses being demonstrated with these vaccines, this project will now apply this technology to therapeutic vaccines. A therapeutic vaccine is one in which the vaccine is used after infection occurs, aiming to induce immunity to alter the course of disease. However, the situation is more complicated when developing therapeutic cancer vaccines. Unlike bacteria and viruses that are generally recognised as foreign to our immune system, cancer cells more closely resemble our normal, healthy cells. Furthermore, tumours are generally unique to the individual and have their own distinguishing antigens. As a result, to develop effective therapeutic cancer vaccines, personalisation of the vaccine is needed. The use of self-amplifying RNA vaccines can address this through the rapid sequencing of the DNA within tumour cells to identify unique cancer peptides. These peptides can in turn be coded within self-amplifying RNA and incorporated into RNA vaccines.

Challenge: The current method for manufacturing lipid nanoparticles is an ethanol injection method; this batch production method involves multi-unit operations (up to 18) with the entire production being carried out in an aseptic area. The complexity is immense and hence scale up and technology transfer is a major challenge. We will now develop this technology to develop therapeutic vaccines for the treatment of cancer and we will investigate formulations to address thermo-instabilities currently associated with SAM-LNP vaccines.

Addressing the Challenge: This project aims to develop thermostable SAM-LNPs which can be manufactured using flexible and adaptive processes, which can be scaled up and down in a rapidly responsive mode. To achieve this, the objectives are: 1) investigate the role of lipid section in controlling vaccine efficacy of SAM-LNP vaccines, 2) format a thermostable vaccine system by exploiting the freeze-drying protocols and 3) develop innovative microfluidic and inline monitoring processes to create continuous manufacturing options for SAM-LNP vaccines.