tebu-bio is a European company specialised in providing innovative reagents and laboratory services in Life Sciences

Need an answer now?

Give us a call

Leave a message, we'll get back to you

CleanCap mRNA research updates

Latest Trilink RNA technology publication.

Thermostable RNA Vaccine Delivery Promises Enhanced Pandemic Response

RNA-based vaccines offer several advantages over traditional vaccines. These include faster production, simpler scale-up, and more rapid adaptability to new targets. Such factors have proven critical in tackling the ongoing COVID-19 pandemic, allowing vaccines targeting SARS-CoV-2 to be developed in record time and enabling vaccination strategies to evolve quickly as new strains of the virus emerge. Both Pfizer/BioNTech and Moderna received regulatory approval for their mRNA vaccine products within a year of the novel coronavirus first being sequenced and many more RNA vaccines are currently in clinical trials.

A main challenge faced by those developing RNA-based vaccines concerns their distribution, particularly at the levels necessary to control a pandemic. This is because RNA-based vaccines are notoriously unstable, being prone to cleavage by ubiquitous ribonucleases. Although RNA engineering has gone some way toward addressing this issue, deep cold chain storage remains essential. The authorized Pfizer/BioNTech and Moderna vaccines require shipping and storage at -70°C and -20°C, respectively. This can be problematic in settings with a well-established medical infrastructure but is especially difficult to implement in regions where resources are more limited.









Lipid nanoparticle (LNP) systems are currently the preferred method for RNA vaccine delivery, and they are being utilized by all SARS-CoV-2 RNA vaccines in clinical trials to date. In LNPs, the negatively-charged hydrophilic RNA is encapsulated by a multicomponent lipid system, which functions to protect the RNA from RNase degradation while enabling its uptake across cell membranes by endocytosis. However, because freeze-thawing impacts the colloidal stability of LNPs, a thermostable RNA vaccine delivery system promises many benefits.

A recent article posted to bioRxiv, a preprint server, describes the development of a thermostable nanostructured lipid carrier (NLC) system for RNA vaccine delivery. Unlike LNPs, NLC particles have an oil core comprised of both solid (trimyristin) and liquid (squalene) lipids, surrounded by surfactants and DOTAP, a cationic lipid. The latter has been widely used for gene transfection applications, where it binds DNA to promote its liposome-based transfer into cells. In the NLC system, DOTAP binds RNA to the outside of NLC particles for cellular delivery in a similar manner.

Testing has demonstrated that liquid NLC particles are stable for at least one year when stored at refrigerated temperatures, with the particle size and component concentrations remaining constant over that time. Additionally, if the stored particles are subsequently complexed with RNA, the NLC system retains the ability to protect the RNA against RNase challenge. These features of NLC particles are extremely valuable, as they allow for stockpiling and rapid complexing with pathogen-specific RNA in the event of a pandemic situation.

In addition to being amenable to stockpiling, NLC particles can be complexed with RNA and lyophilized for long-term storage at ambient temperatures. To demonstrate this, the authors of the publication complexed NLC with secreted alkaline phosphatase (SEAP) RNA prior to lyophilization with sucrose (a common lyoprotectant) and storage at a range of temperatures. When compared to complex stored frozen at -80°C or -20°C, or to freshly formed material, lyophilized NLC/SEAP RNA stored at 4°C or 25°C for 8 months performed similarly in an extensive battery of tests. Alongside these studies, TriLink’s CleanCap® OVA mRNA was complexed with NLCs to determine whether the lyoprotectant concentration could be increased. In combination, the findings illustrate the utility of the NLC system for pandemic preparedness and response.

Featured product: TriLink’s CleanCap® OVA mRNA

Article Reference: “A Thermostable, Flexible RNA Vaccine Delivery Platform for Pandemic Response”. Gerhardt et al, bioRxiv 2021.02.01.429283                                                                                                                                                                                                                                                                

You'll also be interested in...                                                    

Manipulating CD38 for a Novel Immuno-Oncology Therapy

A recent study evaluated a novel CAR-NK therapy for AML using CD38 protein. To investigate the feasibility of a CAR-NK approach for AML, the scientists after reducing the endogenous CD38 expression in NK cells, electroporated a custom-synthesized CD38 CAR mRNA from TriLink BioTechnologies into primary NK cells. These genetically engineered CD38 CAR-NK cells showed reduced propensity for cell fratricide. When co-cultured with bone marrow mononuclear cells from AML patients, the CD38 CAR-NK cells demonstrated enhanced cell killing when compared to mock-electroporated NK cells. These studies employed nine different AML patient samples with varying degrees of CD38 endogenous expression and different AML molecular subtypes....

Article reference: “CD38 knockout natural killer cells expressing an affinity optimized CD38 chimeric antigen receptor successfully target acute myeloid leukemia with reduced effector cell fratricide“ Gurney et al, : Haematologica. 2020 Dec 30

Take a look at the other CleanCap publications

Watch the latest Trilink's webinars and videos on mRNA technlogy

Multiple mRNA capping technologies are available today, each with advantages and disadvantages. In this mRNA Basics webinar, Scott Taylor, MS, Senior Alliance Manager at TriLink, will discuss common mRNA capping strategies and which applications they are best suited for. A live Q&A session will follow the presentation.

The innate immune response is critical to consider when designing your mRNA sequence and manufacturing process. In this mRNA Basics webinar, Cory Smith, Senior Technical Inside Sales Specialist at TriLink, will highlight several strategies for minimizing the innate immune response to mRNA therapeutics. A live Q&A session will follow the presentation.

In celebration of mRNA Day 2020, TriLink proudly presents back-to-back webinars by two pioneers of the mRNA therapeutic platform: Dr. Katalin Karikó (BioNTech) and Dr. Drew Weissman (University of Pennsylvania). Dr. Karikó provides an overview of the history of mRNA therapeutics, while Dr. Weissman offers his perspective on the future of mRNA medicines. A moderated Q&A session follows the presentations.

Don't miss this on tebu-bio's blog!

Want to know more about CleanCap?

Read this article on our blog to learn all about the CleanCap technology.


Download the CleanCap documentation

CleanCap technical brochure

Global CleanCap applications brochure