Standard Operating Procedure for Preparation of Nanoparticles for Gene Delivery
1) Purpose
This SOP outlines the process for preparing nanoparticles specifically designed for gene delivery applications. These nanoparticles are used to carry nucleic acids, such as DNA or RNA, into target cells for therapeutic purposes.
2) Scope
This SOP applies to personnel involved in the preparation and optimization of nanoparticles for gene therapy, including plasmid DNA or siRNA delivery.
3) Responsibilities
- Operators: Responsible for preparing gene-loaded nanoparticles following this SOP.
- QA: Ensures the prepared nanoparticles meet quality, particle size, and gene-loading specifications.
4) Procedure
4.1 Selection of Materials
4.1.1 Cationic Polymers or Lipids
- 4.1.1.1 Select appropriate cationic polymers or lipids (e.g., PEI, chitosan, or liposomes) capable of forming complexes with nucleic acids.
4.2 Nanoparticle Formation
4.2.1 Formation of Complexes
- 4.2.1.1 Mix the cationic polymer or lipid solution with the nucleic acid solution (e.g., plasmid DNA or RNA) to form nanoparticles through electrostatic interactions.
4.3 Gene Loading
4.3.1 Gene Incorporation
- 4.3.1.1 Ensure the nucleic acids are efficiently loaded into the nanoparticles by optimizing the charge ratio between the polymer/lipid and nucleic acid.
4.4 Characterization
4.4.1 Particle Size and Zeta Potential
- 4.4.1.1 Measure the particle size using dynamic light scattering (DLS) and zeta potential to confirm the stability and size of the nanoparticles.
4.5 In Vitro Transfection Efficiency
4.5.1 Transfection Assay
- 4.5.1.1 Perform an in vitro transfection assay on cell cultures to evaluate the gene delivery efficiency of the nanoparticles.
4.6 Storage of Nanoparticles
4.6.1 Storage Conditions
- 4.6.1.1 Store the gene-loaded nanoparticles at 4°C or in a freezer, depending on stability data, to maintain nucleic acid integrity.
5) Abbreviations, if any
- DLS: Dynamic Light Scattering
- PEI: Polyethylenimine
6) Documents, if any
- Gene Delivery Nanoparticle Formulation Logbook
7) References, if any
- Protocols for preparing nanoparticles for gene delivery applications
8) SOP Version
Version 1.0
Annexure
Gene Delivery Nanoparticle Formulation Logbook Template
| Date | Batch Number | Nucleic Acid Type | Particle Size | Zeta Potential | Transfection Efficiency | Operator Initials | QA Initials |
|---|---|---|---|---|---|---|---|
| DD/MM/YYYY | Batch Number | DNA/RNA Type | Size in nm | Zeta Potential (mV) | Pass/Fail | Operator Name | QA Name |