mRNA-LNP Formulation and Encapsulation Service
Overview
The clinical success of mRNA therapeutics is entirely dependent on an effective delivery system that can protect the fragile nucleic acid cargo from nuclease degradation while facilitating efficient endosomal escape. Lipid nanoparticles (LNPs) have emerged as the gold-standard platform, providing a robust architecture for systemic and targeted delivery. CD BioGlyco offers a specialized mRNA-LNP formulation and encapsulation service that transforms high-purity mRNA into potent, stable, and bioavailable nanoparticles. Our platform utilizes advanced microfluidic mixing technologies and proprietary lipid libraries to achieve precise control over particle size, polydispersity, and encapsulation efficiency. By optimizing the critical N/P ratio (nitrogen-to-phosphate) and ionizable lipid pKa, CD BioGlyco ensures that every formulation is engineered for maximal therapeutic impact and minimal systemic toxicity, bridging the gap between molecular design and clinical reality.
Core Technologies
Our formulation platform is built on cutting-edge engineering and lipid chemistry to ensure high-performance delivery:
- Microfluidic Impingement Jet Mixing
We utilize state-of-the-art microfluidic platforms that allow for millisecond-scale mixing of the organic lipid phase and the aqueous mRNA phase. This rapid mixing is essential for generating small, uniform particles with high batch-to-batch reproducibility.
- Ionizable Lipid Engineering
We utilize a library of next-generation ionizable lipids designed with optimized pKa values. These lipids remain neutral at physiological pH to reduce toxicity but become protonated in the endosome to facilitate membrane fusion and cargo release.
- Advanced Analytical Suite
Every LNP batch is characterized using dynamic light scattering (DLS) for size and PDI, RiboGreen assays for encapsulation efficiency, and cryo-electron microscopy (Cryo-EM) to verify morphology and internal structure.
- Automated TFF Systems
We employ tangential flow filtration (TFF) for rapid buffer exchange and concentration. This automated process ensures the removal of residual ethanol and maintains the stability of the LNPs throughout the downstream processing stages.
mRNA-LNP Formulation and Encapsulation: Precision Delivery for Genetic Medicines
The mRNA-LNP formulation and encapsulation service at CD BioGlyco is designed to support researchers from early discovery through pre-clinical development. We offer formulation screening services where multiple lipid ratios or ionizable lipid candidates can be tested in parallel to identify the most potent delivery vehicle for a specific mRNA sequence. Our service scales range from small-volume discovery batches (1-5 mL) to larger pre-clinical batches (500 mL+) suitable for toxicology and non-human primate (NHP) studies. Furthermore, we provide specialized encapsulation for diverse RNA types, including traditional linear mRNA, self-amplifying RNA (saRNA), and circular RNA (circRNA), ensuring that the delivery vehicle is perfectly matched to the unique hydrodynamic properties of the cargo.
Workflow
Formulation Strategy and Lipid Selection
We initiate the process by designing a tailored LNP formulation strategy based on the target tissue and therapeutic application. This involves selecting an optimal combination of an ionizable lipid (for mRNA encapsulation and endosomal release), a structural helper lipid such as DSPC, cholesterol for membrane stability, and a PEG-lipid to control particle size and circulation time, all carefully balanced to achieve the desired delivery performance.
Phase Preparation
The selected lipid mixture is dissolved in a suitable organic solvent, typically ethanol, to form the organic phase. Concurrently, the purified mRNA is prepared in an acidic aqueous buffer, such as citrate or acetate, at a pH that promotes electrostatic interaction with the ionizable lipid. Both phases are independently filtered through 0.22 µm membranes to ensure sterility and the absence of particulates prior to mixing.
Microfluidic Encapsulation
The organic and aqueous phases are precisely pumped into a microfluidic mixing device at defined flow rate ratios (FRR) and total flow rates (TFR). The rapid mixing triggers instantaneous nanoprecipitation, leading to the self-assembly of the mRNA into the lipid core and the formation of uniform, monodisperse LNPs in a reproducible and scalable manner.
Downstream Processing
Immediately after formation, the raw LNP suspension is processed via TFF. This step efficiently removes the organic solvent, performs buffer exchange into a physiologically compatible storage buffer such as PBS or a sucrose-based cryoprotectant solution, and concentrates the LNPs to the target final concentration.
Sterile Filtration and Aliquoting
The concentrated LNP formulation is passed through a 0.22 µm sterile filter to ensure aseptic quality. The sterile product is then aliquoted into low-protein-binding cryovials under a laminar flow hood, ensuring integrity and stability for long-term storage and reducing the risk of contamination or degradation from repeated handling.
Comprehensive Characterization
A final, detailed quality control report is provided for each batch. This includes measurements of particle size and polydispersity index (PDI) via dynamic light scattering (DLS), quantification of encapsulation efficiency (EE%) using a dye-based or enzymatic assay, and assessment of endotoxin levels via the LAL method to confirm suitability for in vivo use.
Publication Data
Journal: Nano Letters
DOI: 10.1021/acs.nanolett.4c06643
IF: 9.1
Published: 2025
Results: In this innovative study, Tanaka and colleagues introduce a streamlined post-encapsulation technique for fabricating mRNA-LNPs by simply incubating mRNA solutions with pre-formed, mRNA-free LNPs at room temperature, bypassing the need for specialized equipment or thermal stress. They elucidate that this efficient encapsulation hinges on a dual-phase mechanism termed "nucleic acid-bridged fusion," where mRNA first adsorbs onto the LNP surface via electrostatic interactions and then triggers particle-to-particle fusion, with both steps requiring precise pH control around 6.0 for simultaneous occurrence. The research systematically evaluates key parameters, revealing that LNP size and lipid composition critically influence loading efficiency, with smaller particles and optimized formulations achieving over 70% encapsulation while maintaining mRNA stability. Validation through in vivo experiments demonstrated potent protein expression and immune responses in mice, comparable to conventional methods, highlighting the method's robustness for therapeutic applications such as vaccines. This approach significantly simplifies mRNA-LNP production, enabling accessible, scalable manufacturing for RNA-based therapeutics and accelerating drug development pipelines.
Applications
Prophylactic Viral Vaccines
Formulate mRNA vaccines with LNPs that provide strong adjuvant effects and efficient antigen presentation. Our LNPs are optimized to trigger both robust antibody and T-cell responses against emerging viral threats.
In Vivo Gene Editing
Support the delivery of CRISPR/Cas9 or base-editing components. Our LNPs ensure that Cas9 mRNA and sgRNA are co-delivered to the same cell, maximizing editing efficiency in the liver or other target organs.
Oncology Immunotherapy
Develop LNP-mRNA formulations encoding tumor-associated antigens or cytokines. These particles can be designed for intratumoral injection or systemic delivery to reprogram the tumor microenvironment and activate anti-tumor immunity.
Cardiovascular Regeneration
Formulate mRNA encoding angiogenic factors for delivery to ischemic cardiac tissue. Our LNPs can be optimized for localized retention and sustained release to promote tissue repair following myocardial infarction.
Advantages
Unrivaled Encapsulation Efficiency
Our microfluidic platforms consistently achieve encapsulation efficiencies of >95%. This minimizes the loss of expensive mRNA cargo and ensures that the final product has a high drug-to-lipid ratio.
Precise Particle Size Control
We precisely tune the diameter of the LNPs (typically 60-100 nm) by adjusting flow parameters. This is critical for controlling biodistribution, as smaller particles are often required for deep tissue penetration or lymphatic uptake.
Optimized Endosomal Escape
By utilizing ionizable lipids with fine-tuned pKa values (typically 6.0-6.5), LNPs maximize the release of mRNA into the cytoplasm. This results in significantly higher protein expression compared to traditional liposomes.
Low Systemic Immunogenicity
Our PEG-lipid selection and lipid ratios are designed to minimize the "accelerated blood clearance" (ABC) effect and reduce the induction of pro-inflammatory cytokines, improving the safety profile for repeated administration.
Frequently Asked Questions
Customer Review
CD BioGlyco's LNP formulation service was a game-changer for our gene editing program. We achieved >98% encapsulation efficiency, and the editing rates in our NHP models were significantly higher than with our previous supplier.
— By Dr. H.W., Senior Scientist
The consistency of the LNPs from CD BioGlyco is remarkable. Our batch-to-batch PDI is always below 0.08, which has given our team immense confidence in our in vivo dosing studies.
— By Manager, Vaccine Development
We were struggling with the stability of our large saRNA cargo. CD BioGlyco optimized a custom LNP formulation that kept the RNA intact and delivered high protein expression in our mouse models.
— By Dr. C.R., Associate Professor
Associated Services
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CD BioGlyco provides the engineering precision and lipid expertise required to bring your mRNA therapeutic to life. Our mRNA-LNP formulation and encapsulation service is the bridge between genetic design and clinical efficacy. Let us help you navigate the complexities of delivery to ensure your project reaches its full therapeutic potential, contact us!
Reference
- Tanaka, H.; et al. A post-encapsulation method for the preparation of mRNA-LNPs via the nucleic acid-bridged fusion of mRNA-free LNPs. Nano Letters. 2025, 25(16): 6445-6453. (Open Access)