Standard Operating Procedure for Preparation of Nanoparticles for Gene Delivery
1) Purpose
This SOP outlines the procedure for preparing nanoparticles intended for gene delivery. Nanoparticles are used to deliver nucleic acids (e.g., DNA, RNA) to cells, protecting the genetic material and enhancing cellular uptake for gene therapy applications.
2) Scope
This SOP applies to personnel involved in the formulation and characterization of nanoparticles designed for gene delivery in pharmaceutical or genetic research settings.
3) Responsibilities
- Operators: Responsible for preparing gene delivery nanoparticles as per the outlined procedure.
- QA: Ensures the nanoparticles meet the required specifications for nucleic acid encapsulation, particle size, and gene transfection efficiency.
4) Procedure
4.1 Selection of Nucleic Acids and Nanoparticles
4.1.1 Nucleic Acid Selection
- 4.1.1.1 Choose the appropriate nucleic acid (e.g., plasmid DNA, siRNA, mRNA) based on the intended gene therapy application and ensure its stability during nanoparticle formulation.
4.1.2 Nanoparticle Material Selection
- 4.1.2.1 Select biodegradable and biocompatible polymers (e.g., chitosan, PLGA) or lipids for nanoparticle formation, optimizing for nucleic acid protection and cellular uptake.
4.2 Nucleic Acid Incorporation
4.2.1 Encapsulation or Complexation
- 4.2.1.1 Encapsulate or complex the nucleic acid with the nanoparticles using electrostatic interactions or entrapment within the nanoparticle matrix. Optimize for high encapsulation efficiency while maintaining nucleic acid activity.
4.3 Characterization
4.3.1 Particle Size and Encapsulation Efficiency
- 4.3.1.1 Measure the particle size using dynamic light scattering (DLS) and evaluate encapsulation efficiency using gel electrophoresis or spectrophotometry to ensure proper formulation.
4.3.2 Gene Transfection Efficiency
- 4.3.2.1 Perform in vitro transfection studies using target cells to assess the ability of the nanoparticles to successfully deliver the genetic material and express the gene of interest.
4.4 Stability and Storage
4.4.1 Storage Conditions
- 4.4.1.1 Store the gene delivery nanoparticles in sterile containers at 4°C or -20°C depending on stability data, ensuring the integrity of the nucleic acids is preserved over time.
5) Abbreviations, if any
- DLS: Dynamic Light Scattering
- PLGA: Poly(lactic-co-glycolic acid)
- siRNA: Small Interfering RNA
6) Documents, if any
- Gene Delivery Nanoparticle Formulation Logbook
7) References, if any
- Protocols for nanoparticle preparation for gene delivery
8) SOP Version
Version 1.0
Annexure
Gene Delivery Nanoparticle Formulation Logbook Template
| Date | Batch Number | Nucleic Acid | Nanoparticle Material | Particle Size | Encapsulation Efficiency | Transfection Efficiency | Operator Initials | QA Initials |
|---|---|---|---|---|---|---|---|---|
| DD/MM/YYYY | Batch Number | Nucleic Acid Used | Material | Size in nm | Efficiency (%) | Transfection (%) | Operator Name | QA Name |