LNP Versatility

One of the greatest strengths of LNPs is their versatility. However, this comes with a price - LNP formulations do not conform to a one-size-fits-all approach. 

The versatility of LNPs is achievable in part through the diversity of lipids available for these applications. Each lipid has a set of unique biophysical properties, which impacts the structural and functional properties of the LNP. Tailoring the lipid composition of the LNP can be used to adjust tissue- and/or cell-specific targeting, uptake, immunogenicity, and biocompatibility.¹ However, an optimized LNP for one scenario may not translate to a different target and therapeutic payload. Everything needs to be tuned for the target and indication in mind. 

It is important to have access to lipid synthesis expertise and a large library of research-ready lipids to maximize the opportunity to find an optimal formulation and benchmark against published formulations when seeking synthesis of novel lipid components or exploring novel lipid formulations. 

Lipid synthesis is complex, requiring specialized expertise, analytical equipment, and facilities to produce and characterize pure, high-quality lipids. The use of high-quality lipids in LNP formulations is of utmost importance. Lipids are easily oxidized, and oxidized lipids can form lipid-mRNA adducts, which reduce the activity of the LNP formulations.² To prevent this misfortune, lipids used to develop LNPs must be prepared, handled, and stored with care at all stages of LNP preparation and development. 

LNP Preparation

Microfluidic mixers are the gold standard for LNP production. They offer precise control over LNP formulation parameters, ensuring reproducibility of the final preparation and the ability to rapidly produce different LNPs under variable processing parameters and experimental conditions to find the optimal formulation.³

However, microfluidic mixing devices can require significant initial investment which limits their accessibility. Some researchers may choose to use lower-cost options, but these methods may have limited reproducibility and difficulty with scalability in the final formulation.

Without proper handling, lipids can be fickle to solubilize, and RNA is prone to degradation. Loss in integrity of either of these components can undermine the potency of the final formulation. Solvent removal and sterile filtration are critical post-processing steps that help ensure that the LNPs are homogenous, stable during storage and use, and free from any residual chemical or biological contaminants. 

LNP Characterization & Screening 

LNP formulation, characterization, and screening requires a broad range of analytical methods that are performed as part of LNP optimization. Given the sheer number of parameters to adjust, it is possible that there will be multiple LNP formulations produced during an LNP screening experiment. It is then paramount to identify a small subset to further qualify with  in vivo studies. 

Although  in vitro studies do not always correlate with  in vivo efficacy, this is not the rule.^4,5 Several candidate LNP formulations have been identified with  in vitroscreening methods that subsequently performed well in  in vivo experiments.^6-9
Well-designed in vitro LNP screening can be used as a selection tool to narrow down an otherwise large number of LNP formulations to those that are the least risky. 

These methods can be made more robust by including benchmark LNP formulations that are well-known for their performance in each context, as well as by using multiple biological endpoints and several cell culture models to increase confidence. Taken together,  in vitro screening can identify early lead candidates appropriate for follow-up studies with  in vivo models or produce proof-of-concept data critical for securing early funding. 

Preparing for GMP

Once an optimal formulation is identified, additional challenges remain to translate the formulation into the scale required for final GMP formulation and ultimately, clinical trials and acceptance for use in humans. Microfluidic devices or other small-scale fluid-injection based devices may not have directly translatable equivalents for larger scale manufacturing, requiring fine-tuning of formulation and manufacturing at liter to multi-liter scale. Likewise, GMP manufacturing of LNP components, particularly if novel lipids are a part of the final formulation, will be required as part of the final method transfer to an accredited CDMO. Additional time (months to >1 year) are likely to be associated with such GMP formulation development, and significant licensing fees may be associated with use of patented third-party formulations.   

From R&D to GMP, Cayman Supports LNP Development 

With the breakneck speed at which LNPs are revolutionizing modern medicine, the competition in this space is fierce. Given the high costs of equipment and resources required to develop LNPs as well as a murky intellectual property climate, researchers and biotechnology startups in the pharmaceutical industry are seeking to outsource their LNP R&D to service providers equipped with the expertise and facilities needed for this undertaking. 

But finding an R&D partner is not as straightforward as it seems. Experiments performed during R&D set the course of further LNP development. Early R&D projects will benefit from choosing a service provider that has the flexibility to adapt to changes in project direction, and with in-house capabilities to support your project not only with early formulation and characterization, but also with lipid expertise and in vitro, assay development, and screening capabilities to cater to your specific project aims. 

Cayman supports our clients by providing tailored solutions for LNPs, whether you require single-point or end-to-end support. We work with you to develop the best experimental design within your budget constraints to provide high-quality data with prompt, professional communication and quick turnaround.

Backed by 40 years of lipid expertise and an interdisciplinary team of scientists, Cayman offers an industry-leading collection of ready-to-use lipids and research-ready LNPs, as well as on-site R&D facilities for LNP development, characterization, and screening. Our experts can also help design and synthesize novel ionizable lipids to expand your intellectual property portfolio, and they have the foresight and in-house process development expertise to guide you toward scalable, GMP-compatible components for production in GMP suites at our Ann Arbor, Michigan headquarters. Cayman will work closely with your preferred formulation partner to ensure a smooth transition of identified formulations and any custom components to advance your program toward clinical trials.