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LNP Profiler Kit with AutoLNP
Your all-in-one solution for LNP characterization
Key features
Robust
A reliable sample-to-answer workflow to visualize and quantify sample populations and single LNP for size, cargo loading, and ligand positivity and abundance
Optimized
Characterize different PEG-based LNP formulations with nanometer precision to better understand uptake efficacy, and evaluate integrity over time
Simplified
Powered by AutoLNP for automated benchtop imaging and analysis, delivering cost-effectiveness and throughput, with minimal training needed
Rapid
Analyze tens of thousands of particles in one day and instantly obtain reproducible data to accelerate LNP development and quality testing
Application Kit™
LNP Profiler with AutoLNP
The LNP Profiler Kit with AutoLNP is the first verified solution for characterizing lipid nanoparticles (LNPs) using super-resolution microscopy. It delivers rapid, high-resolution, quantitative results for profiling LNP formulations down to individual particles, using optimized reagents to capture PEG-containing LNPs and label them with PanLNP, RNA cargo and surface ligand stains. This allows for detailed visualization and quantification of LNP size (< 3 nm SD between chips), integrity, cargo loaded fraction, and ligand abundance with unmatched sensitivity (< 5% SD between chips).
Combined with AutoLNP for automated imaging and analysis, researchers can gain instant, valuable insights to accelerate LNP formulation optimization, and improve manufacturing output. This all-in-one benchtop assay is reliable, faster and more cost-effective than CryoEM, and offers higher resolution than confocal microscopy, delivering a new standard for LNP analysis to accelerate next-gen therapeutics.
What’s in the kit?
-
- Surface Block
- LNP Surface Reagent
- Anti-PEG Capture
- LNP Wash Buffer
- Four LNP Assay Chips, with 4 lanes each
- Pan-LNP Detection (640)
- Human IgG Ligand Detection (560)
- Cargo Detection (488)
Note: The LNP Profiler kit has a shelf-life of three months. AutoLNP™ requires a CODI account and an active internet connection. Offline support planned for a future release.
LNP Characterization and Profiling Redefined
Determine size & morphology
Size LNPs, assess sample morphology and integrity
LNP Profiler kit empowers researchers to optimize LNP formulations with super-resolution analysis of populations down to single particles, using specific capture on passivated chips and optimal labeling.
Key outputs:
- Morphological visualization of nanoparticles using PanLNP stain, Aplo Scope, the Nanoimager, AutoLNP acquisition app and CODI software. With final reporting including the overall LNP size distribution per lane and Polydispersity Index (PDI).
- Super-resolution sizing with nanometer precision using machine-learning algorithms, with the SD of LNP diameter being < 3 nm from chip to chip.
- Quick evaluation of sample integrity, stability, aggregation or fragmentation
- Faster sample to answer results than with CryoEM or TEM
Assess cargo encapsulation
Determine cargo positivity and loaded fraction
The LNP Profiler kit includes a nucleic acid stain for diffraction-limited imaging to evaluate cargo encapsulation per particle alongside super-resolution imaging of PanLNP and ligand stains.
Key outputs:
- Real-time information about LNP count in minutes, with simultaneous analysis of cargo encapsulation performed with single LNP resolution using AutoLNP and CODI
- Increased sensitivity to determine cargo loaded fraction per sample and cargo positivity with SD of < 5% from chip to chip
- Valuable quantitative information from LNP formulations with plots displaying relative cargo (photon count from diffraction-limited imaging) and ligand abundance (localizations per single LNP) using a threshold for ligand positivity.
Quantify ligand loading
Evaluate LNP ligand engineering
Assess engineering processes and analyse functionalized particles through the detection and quantification of surface ligands using single-molecule super-resolution microscopy.
Key outputs:
- Quantify LNP ligand engineering down to individual particles, with SD of ligand positivity being < 5% from chip to chip
- Measure particle size, cargo encapsulation and ligand abundance and loading efficiency in one assay using diffraction-limited and super-resolution microscopy for rapid, highly precise results
- Streamlined sample-to-answer workflow that allows visualization and analysis of tens of thousands of particles per experiment, with reduced hands-on time and minimal training required
Correlate LNP metrics
Multiple LNP metrics in a single, high-precision assay
Analyze LNP formulations in depth with individual particle images and LNP metric correlation.
Key outputs:
- Use morphology and size range of individual LNPs to gain insights into particle uptake and delivery based on ligand abundance and cargo positivity.
- Preview snapshot images of single LNPs and gain valuable insights into LNP formulations with a detailed two-page report.
- Correlate cargo-positive LNPs with surface ligand concentrations, using plots displaying relative cargo within each lane (photon count from diffraction-limited imaging) against ligand abundance (localizations per single LNP), LNP size distribution as a function of positivity, and LNP size distribution as a function of cargo loading.
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AutoLNP™: LNP acquisition, analysis, and reporting software
Experiment setup
Experiment setup
The first step of AutoLNP is to input your experiment settings, such as the experiment name and collaboration and any relevant tags. Optimized acquisition and analysis settings for the LNP Profiler kit are set as default for users, including channel names, laser power, number of frames, exposure time, and optimal analysis workflow for automated and accurate end to end workflow. Settings can be adjusted to fit different experimental settings, easily saved and applied across all lanes.
One-click channel mapping
One-click channel mapping
Channel mapping is an essential step towards generating accurate multicolor super-resolution data using a calibration slide. AutoLNP makes channel mapping even easier with a one-click procedure that only takes around 2 minutes using a bead slide. The channel mapping tool optimizes the 560 nm and 640 nm laser powers, acquiring as many fields of view as necessary to complete the calibration.
Calibration & Sample Check
Calibration & Sample check
System calibration is a critical step that uses both Autofocus and AutoTIRF tools to automatically find the imaging surface of the LNP Profiler Assay Chip, lock the focus on the chip, and determine the optimal TIRF illumination angle for higher imaging quality. Sample pre-check is an optional tool that quickly scans each lane of the chip, providing representative images of each lane. This helps confirm sample calibration, optimal focus and illumination, as well as sample density and particle staining.
Field of view selection
Field of view selection
Field-Of-View (FOV) selection can be performed either by choosing the number of FOVs to be imaged at the top of each lane, or manually selecting FOVs within each lane’s imaging area. The tool includes estimated time indicators to predict experiment duration and a color-coded system to easily visualize the position of the currently selected FOV, those already acquired, or those used for calibration to avoid repeated imaging. On a Nanoimager microscope, AutoLNP images a FOV of 50 µm by 80 µm, with each imaging area surrounded by a bleaching safe zone so that neighboring FOV are not affected by stray laser light.
Automatic 4-lane acquisition
Automatic 4-lane acquisition
A live camera view shows blinking fluorophores during super-resolution imaging. The software provides simultaneous real-time image analysis and information including live localizations from the blinking fluorophores and overall experiment progress. After being acquired, each dataset is automatically uploaded to CODI, thus, AutoLNP also provides information on the FOVs being imaged, those being uploaded to CODI, those being analyzed, and the FOVs where analysis is completed and the number of particles/clusters found.
Summary, LNP montage & Data export
Summary, LNP Montage & Data export
The acquisition summary page offers a visual overview of acquired datasets, showcasing imaging settings, FOVs used either for imaging or calibration, links to analyzed datasets on CODI, and the clusters counts for each analysis. It also displays a mosaic of randomly selected LNPs, representative of each positivity class. Users can export analyzed datasets using three tools: the LNP montage tool for snapshots of LNP clusters grouped by signal positivity and size; a two-page report summarizing LNP population details including counts per lane and FOV, size distribution, Polydispersity Index (PDI), biomarker positivity per lane, LNP size distribution per sample as a function of positivity, and relative cargo and ligand abundance; and a CSV file with all LNP data for downloading and sharing.
Note: image above shows a collage of the different data outputs from AutoLNP and CODI.
FAQs
How long does it take to run the LNP Profiler assay end-to-end?
The LNP Profiler kit with AutoLNP has been designed to provide fast, actionable data on LNP formulations. 16 LNP samples can be captured, stained, imaged and analyzed in under 8 hours. In addition, the entire workflow has been simplified so that users of any skill level can characterize LNP formulations with super-resolution microscopy with minimal training.
Is the kit compatible with third-party antibodies and slides?
AutoLNP is built to automatically acquire field of view in the lanes of the 4-lane LNP Profiler Assay Chip with a single click, and is not intended to be used with any other slides or coverslips. The LNP Profiler kit provides all necessary reagents to detect and size PEG-based LNPs using PanLNP and surface antibody ligand stains (in the 640 and 561 channels, respectively) using SMLM. The 488 channel is used for cargo detection in diffraction-limited mode. Users can swap these reagents for their own antibodies/dyes, however, thorough optimization would be required as the kit reagents might not work and different software tools might be needed for accurate analysis.
Does AutoLNP have manual controls as well?
AutoLNP introduces great new algorithms like AutoFocus, AutoTIRF, and automated acquisition across lanes. You can manually set the illumination (TIRF) angle if desired, and you can adjust the focus on a per-lane basis prior to starting the automated acquisition, so you can be sure to have the imaging results you want, all while being able to step away from the microscope for imaging.
What if my LNPs do not contain PEG or use human antibody ligands?
For non-PEG based LNPs, this assay will not capture or detect particles with this current design but please reach out as our application teams can support with possible alternative options. Additionally, development for future ligand, capture, and surface detection methods are ongoing. Similarly if your LNPs use a different ligand type, a new detection reagent could be swapped into the 560 channel but testing and optimization would be required. Super-resolution will still be superior to alternative characterization methods and we would be happy to discuss how we can help with your unique application or LNP type.