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