In this study, we formulated LNPs with SARS-CoV-2 Spike mRNA as a model antigen. The LNPs were based on established ionizable lipids, and some included putative adjuvants as additional components of the LNPs. A cohort of mice was immunized, and immune responses were assessed by ELISA for total and neutralizing antibody production, flow cytometry for T cell subsets, and ELIspot for Spike peptide-specific immune responses. We found that the different LNP formulations stimulated different aspects of the immune response, which can inform future approaches to vaccine development.

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To cite this poster: Rumble, J.M., Gronevelt, J.P., Blanks, A., et al. ​Lipid nanoparticle (LNP) components dictate the cellular immune response to vaccination. Poster presented at: The Annual Meeting of the American Association of Immunologists; May 3-7, 2025. Honolulu, HI. 

In this study, LNPs encoding SARS-CoV-2 Spike protein were formulated with several different lipids proposed to stimulate immune responses. These LNPs were used to transfect human monocyte-derived dendritic cells (MDDCs), and Spike-derived MHC class I and class II peptides were identified. Evaluation of immunopeptidomic differences between these formulations as well as understanding of resulting immune responses will help the design of vaccine LNPs in the future.

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To cite this poster: Rumble, J.M., Gronevelt, J.P., Nyayapathy, S., et al. ​Lipid nanoparticle SARS-CoV-2 vaccine-mediated immunopeptidome identification. Poster presented at: The Annual Meeting of the American Association of Immunologists; May 3-7, 2025. Honolulu, HI. 

In this study, we investigated the expression of three different nucleic acid cargo types (mCherry mRNA, GFP DNA and Cy5-labeled 2′3′-cGAMP) delivered to three different ovarian cancer cell lines via four loadable LNPs containing benchmark lipids SM-102, ALC-0315, (S)-C12-200, or DLin-MC3. We then compared the relative efficacy of each formulation in delivering each payload to the various ovarian cancer cells (PA-1, SK-OV-3, and Caov-3) cultured as monolayers vs spheroids to assess whether the optimal LNP formulation is consistent across 2D and 3D culture models.

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To cite this poster: Taylor, D., Forsyth, V., Riddering, C., et al. ​Lipid nanoparticle screening in 2D vs 3D cultures: Ovarian cancer cell lines exhibit different preference for optimal lipid formulation in monolayers vs spheroids. Poster presented at: American Association for Cancer Research Annual Meeting 2025; April 25-30, 2025; Chicago, IL. 

CAR-T cell therapy is a groundbreaking form of immunotherapy, with seven US FDA-approved therapies for hematological malignancies and ongoing investigations for solid tumors. This autologous therapy includes harvesting patient’s T cells, engineering them to express CAR constructs, and transfusing them back into the patient to target and destroy cancer cells. In this study, we evaluated five LNP formulations to express a reporter gene (eGFP mRNA) in microbubble-CD3/CD28-activated T cells and stimulated with IL-2 and IL-7.

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To cite this poster: Nyayapathy, S., Gronevelt, J.P., Holcomb, E., et al. ​Lipid nanoparticle-mediated in vitro transfection of microbubble-activated T cells. Poster presented at: American Association for Cancer Research Annual Meeting 2025; April 25-30, 2025; Chicago, IL. 

Key Features

• Cayman’s LNP development services in partnership with Labcorp utilizes in vivo imaging technology to create a comprehensive LNP screening platform. 
  
• With this approach, three LNP formulations were assessed in vitro and in vivo for potency of reporter gene expression and differential organ delivery. 
  
• This comprehensive LNP formulation, characterization, and analysis service can be used to identify candidate LNP formulations with tissue-tropic distribution. 
  

In this study, we investigated the efficiency of LNPs loaded with GFP-encoding pDNA using several previously published ionizable lipids, including DLin-MC3-DMA (MC3), CIN-16645 (LP-01), ALC-0315, 4A3-SC8, and DOTAP (SORT), SM-102, and β-sitosterol as a cholesterol analogue in SM-102. Key formulation characteristics, such as polydispersity index (PDI), Z-average diameter (Z-Avg), encapsulation efficiency (%EE), and freeze-thaw stability, were examined through plate-based methods. Our results show that all LNP formulations achieved ≥85% encapsulation of pDNA-eGFP with PDI values ≤0.10, indicating efficient nanoparticle formation. Next, we evaluated the transfection efficiency and mean fluorescence intensity (MFI) of pDNA-loaded LNPs in Huh-7 hepatocytes and A549 lung epithelial cells. We show that pDNA-eGFP-loaded LNPs exhibited comparable GFP intensities, with slightly lower transfection efficiencies than mRNA-loaded LNPs. Both Huh-7 and A549 cells were successfully transfected with pDNA-encapsulated LNPs

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To cite this poster: Konopka, S., Stewart, S., Gronevelt, J.P., et al. In vivo investigation of pDNA delivery using lipid nanoparticle formulations in A549 and Huh7 cell lines. Poster presented at: Cell Bio 2024; December 14-18, San Diego, CA. 

Ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) suppresses innate immunity and promote tumor progression and metastasis in the tumor microenvironment. ENPP1 inhibits STING signaling by hydrolyzing the degradation of 2',3'-cGAMP, blunting induction of the type 1 interferon response and antitumor immunity.

Listen as David Taylor, Ph.D., a scientist in Cayman's Assay Research & Development group, modulates ENPP1 expression and STING activation in ovarian cancer cell lines and macrophages with LipidLaunch™ Loadable LNPs and explores the contribution of ovarian cancer cell-derived extracellular vesicles in the tumor microenvironment.

Presented as part of the 2024 Tumor Microenvironment and Therapy Symposium at The University of Michigan.

Read the Webinar Highlights for a quick overview of the key takeaways. 

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Learn more about LipidLaunch™ LNPs and reagent kits.

Key Features

• Primary immune cells, especially T cells, are challenging to transfect due to their natural resistance to foreign genetic material.
• Lipid nanoparticles (LNPs) offer improved efficiency with reduced cytotoxicity compared with traditional transfection methods.
• This workflow can be used with LNPs for the delivery of mRNA to achieve optimal protein expression in activated primary human T cells.

Beyond its implications in disease, apoliporotein E (ApoE) has been implicated in the intricate process of lipid nanoparticle (LNP) uptake. Recent studies have revealed the essential role of ApoE in facilitating the cellular internalization of LNPs, providing a potential avenue for targeted drug delivery and therapeutic interventions. In this study we have expressed and purified all three ApoE isoforms and characterized their binding properties to cognate ligands by surface plasmon resonance (SPR). Furthermore, we show that Cayman-produced LNP uptake is enhanced by addition of exogenous ApoE in a cell-based assay with lung epithelial cells. 

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To cite this poster: Guerra, A.J., Muzzarelli, K.M., Gronevelt, J.P., et al. Apolipoprotein E: Purification, characterization, and lipid nanoparticle uptake enhancement. Poster presented at: PEGS Boston Summit; May 13-17, 2024. Boston, MA.

Key Features

• Cayman’s LipidLaunch™ SM-102 LNP Kit (Loadable) is comprised of lyophilized SM-102-based lipid nanoparticles (LNPs) prepared without cargo.
• Allows researchers to encapsulate RNA cargo of choice for LNP-mediated transfection without microfluidic mixing devices.
• Enable RNA delivery to difficult-to-transfect cell lines and primary cultures.
• LipidLaunch™ SM-102 LNPs (Loadable) facilitate effective RNA transfection with minimal cytotoxicity compared to traditional transfection reagents.
  

Lipid nanoparticles (LNPs) represent an extremely promising technology for delivery of gene products for a host of potential health applications, from immunization against emerging pathogens to personalized neoantigen cancer therapy and gene therapy for many diseases. Combining our industry-leading expertise in lipid chemistry, synthesis, and analysis with proficiencies in bioanalysis and cell biology, Cayman offers comprehensive services to support LNP development, characterization, and screening.

Key Features

• Cell lines derived from different tissues have distinct reporter expression patterns in response to a variety of lipid nanoparticles (LNPs).
  
• Serum requirement for LNP uptake and cargo expression in vitro vary with cell type and ionizable lipid.
• Experimental design is of utmost importance in assessing the efficacy of any given LNP preparation.
  

Inflammation is the immune system's response to potential danger signals and damage. The coordination of immune cells, inflammatory lipid mediators, and cytokines act to mount the appropriate immune response, resolve inflammation, and restore homeostasis. Immunomodulation is a powerful strategy that can be used to revolutionize the treatment of autoimmune diseases, cancer, and countless other inflammation-related disorders. Cayman offers a wide range of tools to help you study the factors related to innate and adaptive immune responses.

​Listen as Julie Rumble, PhD, Director of Immunology Services at Cayman Chemical, discusses the workflow for formulation, analysis, and testing of LNPs in multiple cell types. 

Presented by Unchained Labs and Fierce Biotech. 

Do you have a question or comment for Dr. Rumble? Let us know.