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Lipid Nanoparticles

Lipid nanoparticles (LNPs) stand as the most advanced non-viral gene delivery system within clinical practice. They have proven their capability to safely and efficiently transport nucleic acids, addressing a significant obstacle that previously hindered the progress and utilization of genetic medicines.

Genetic medicine encompasses various applications, including gene editing, the expedited development of vaccines, immuno-oncology, and the treatment of rare genetic and previously untreatable diseases. All of these applications have traditionally faced challenges due to inefficiencies in nucleic acid delivery.

 
Multiple Lipid nanoparticle mRNA vaccine, 3D illustration
Lipid Nanoparticles are a class of nanoscale delivery systems designed to transport and protect therapeutic molecules, such as drugs and RNA, to specific target sites within the body. They have gained significant attention in the field of medicine and biotechnology due to their ability to enhance the bioavailability and efficacy of drugs by improving stability and ability to target points of interest for drug delivery.
 
The basic structure of an LNP consists of a lipid bilayer surrounding a hydrophobic core. This structure allows the LNPs to encapsulate hydrophobic drugs or nucleic acids within the core while keeping the hydrophilic components on the surface, making them stable and compatible with the aqueous environment of the body.
lipid nanoparticle structure

Modern LNPs comprise four essential components: cationic/ionizable lipid, helper lipid, cholesterol and PEGylated lipid. Each serves specific functions in the delivery system (Fig. 2).

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Essential components of LNPs

LNP Introduction 

Introduction
Working Principle

Working Principle

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LNP Synthesis & Formulation 

Conventionally, solvent injection and thin film rehydration are two common methods for liposome synthesis. Due to the simplicity of equipment requirements, such as sonicators or rotary evaporators, solvent injection and thin film rehydration methods are widely adopted in both research and production process. However, these traditional methods meet the challenges of low homogeneity of resulted LNPs, hash processes for delicate biomolecules, such as DNAs, RNAs or proteins. 
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Overview of LNP Synthesis, Formulation and Manufacturing Methods

This report reviews the various traditional approaches to LNP synthesis, contrasts them with microfluidic synthesis, and provides an overview of key synthesis parameters and downstream processing methods. While several traditional manufacturing methods have laid the foundation for current LNP technologies, modern methods use microfluidics for superior precision, reproducibility, and scalability. 

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Introduction to Lipid Nanoparticle: LNP Formulation Design and Preparation 

This technical introduction explores lipid nanoparticle types, the formulation design, and preparation workflow of LNPs, highlighting common types, their typical composition, supported cargoes, and basic preparation considerations.

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Targeted Delivery of Lipid Nanoparticles Through Surface Modification 

This report explains the structural basis of LNP targeting, passive and active targeting mechanisms, and cutting-edge surface engineering techniques. Surface modifications— ranging from ligand conjugation to advanced lipid engineering — play a critical role in overcoming biological barriers, enhancing cellular uptake, and directing LNPs to specific tissues or cell populations. 

To address these challenges, microfluidic mixing method has been rapidly developed in the past decade.
Slide1_ Slide1 how LNP synthesized

Schematic of oligonucleotide based LNP synthesis

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LNP/liposome preparation protocol

The preparation protocol is illustrated by using NanoGenerator instrument.

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Lipid Nanoparticles / Liposome Synthesis 
by NanoGenerator™ Microfluidic Mixing System

Technical note for Lipid Nanoparticles / Liposome Synthesis by NanoGenerator™ Microfluidic Mixing System 

Small volume(0.1-0.5 ml) lipid nanoparticle preparation for drug discovery and screening

LNP formulation tutorial with 1-4 samples per run

Automated high throughput screening LNP platform

Media volume(1- 200 ml) lipid nanoparticle process for formulation development and optimization, candidate selection in the pre-clinical study

Large volume (>100mL) lipid nanoparticle process for clinical development, GMP manufacturing.

Preparation Protocol

Payloads

lipid nanoparticle synthesis mRNA

mRNA

peptide lipid nanoparticle synthesis

Proteins and peptides

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circRNA

dna lipid nanoparticle synthesis

DNA

small molecule lipid nanoparticle synthesis

Small molecules

Applications

Gene Therapy

lnp for gene therapy

Genetic drugs, including small interfering RNA (siRNA), mRNA, or plasmid DNA, hold promise for treating a wide array of diseases by either suppressing harmful genes, producing therapeutic proteins, or employing gene-editing techniques. Currently, lipid nanoparticle (LNP) systems stand at the forefront among non-viral delivery methods, enabling the clinical utilization of genetic drugs.

Vaccines

LNP for vaccine

Lipid nanoparticle has played a pivotal role in expediting the development of vaccines, as demonstrated by Pfizer and Moderna in the COVID-19 pandemic response. It allows for precise and high-throughput screening of potential vaccine candidates. The technology facilitates rapid testing, antigen formulation, and optimization of vaccine delivery systems.

Cell Therapy

lipid nanoparticle for cell therapy

Cell Therapies entail the transformation of cells, commonly immune cells, obtained either from the patient (autologous) or a compatible donor (allogeneic). These modified cells undergo isolation, amplification, and are later reintroduced into the patient. Lipid Nanoparticles (LNPs) present a versatile approach to cell reprogramming, facilitating the transfer of RNA responsible for protein expression or gene editing. 

Other Areas

lipid nanoparticle for liposome cosmetics

Other applications including cosmetics, medical imaging, nutrition, agrochemicals, etc.
The cosmetics sector stood at the forefront in acknowledging and utilizing nanotechnology advancements in diverse product innovations. Liposomal cosmetic formulations are expected to offer several benefits, including improved stability and effectiveness, along with successful ingredient penetration into the skin. A variety of liposomal cosmetics are currently in use.

Application Webinars:

NanoGenerator LNP Synthesis & Application Webinar with ProMab Biotechnologies

Lipid nanoparticle LNP application in gene editing, CRISPR-Cas9

Resources

Publications

Lipid Nanoparticle and Liposome Synthesis

Application

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Precise lipid NP and liposome synthesis by microfluidic mixing methods. Efficient method for generating nanoparticles for targeted drug or nutrient delivery.

Vaccines 2023
 

Publication

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A Single-Dose Intramuscular Immunization of Pigs with Lipid Nanoparticle DNA Vaccines Based on the Hemagglutinin Antigen Confers Complete Protection against Challenge Infection with the Homologous Influenza Virus Strain

Lipid Nanoparticle & Liposome Introduction

Article

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Introduction of lipid nanoparticle and  liposome. It introduces the structure of LNP, LNP formulation, and LNP preparation protocol, etc.

International Journal of Pharmaceutics 2022

Publication

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Lipid nano-vesicles for hormone encapsulation: A comparison between different fabrication technologies, drug loading, and an in vitro delivery to human tendon stem/progenitor cells in 2D and 3D culture

PLGA Nanoparticle Synthesis by NanoGenerator

Application

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Drug-loaded PLGA nanoparticles (NPs) can be synthesized by our system. PLGA NPs synthesis in continuous flow microfluidics by the nanoprecipitation process.

Int. J. Mol. Sci. 2023
 

Publication

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CAR-NK Cells Generated with mRNA-LNPs Kill Tumor Target Cells In Vitro and In Vivo

Industry News

Lipid Nanoparticle Drug Delivery Advances

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It highlights recent advancements in lipid nanoparticle (LNP) drug delivery. Key updates include a new machine learning model from Johns Hopkins for predicting LNP efficiency, a Nature study on safer LNP-based DNA delivery through innovative design, and corporate news. It is the curated source of scientific breakthroughs and industry developments, emphasizing progress in LNP applications for drug and gene delivery, as well as notable shifts within leading biotech companies in this sector.

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