Standard Operating Procedure for Preparation of Nanoparticles for Targeted Drug Delivery

1) Purpose

The purpose of this SOP is to outline the procedure for preparing nanoparticles designed for targeted drug delivery. These nanoparticles are engineered to deliver therapeutic agents directly to specific cells or tissues, thereby enhancing drug efficacy and minimizing off-target effects.

2) Scope

This SOP applies to personnel involved in the formulation and characterization of nanoparticles for targeted drug delivery, particularly in pharmaceutical development and clinical research settings.

3) Responsibilities

• Operators: Responsible for following the outlined procedures for the preparation and characterization of the nanoparticles.
• QA: Ensures that the nanoparticle formulations meet the required specifications for particle size, drug loading, targeting efficacy, and sterility.

4) Procedure

4.1 Selection of Materials

4.1.1 Nanoparticle Material

• 4.1.1.1 Select appropriate materials such as PLGA (poly(lactic-co-glycolic acid)), PEGylated lipids, or other biodegradable polymers that provide stability and facilitate targeted delivery.

4.1.2 Drug and Targeting Ligand

• 4.1.2.1 Select the therapeutic agent based on its suitability for targeted delivery (e.g., chemotherapeutic agents, peptides).
• 4.1.2.2 Choose targeting ligands (e.g., antibodies, peptides) specific to receptors on the target cells or tissues.

4.2 Nanoparticle Preparation

4.2.1 Nanoparticle Synthesis

• 4.2.1.1 Use solvent evaporation, nanoprecipitation, or emulsification methods to prepare nanoparticles. Optimize the process to ensure a particle size between 100-200 nm for effective targeting.

4.2.2 Surface Functionalization

• 4.2.2.1 Conjugate targeting ligands (e.g., antibodies, aptamers) to the surface of the nanoparticles using techniques such as carbodiimide cross-linking (EDC/NHS) or thiol-maleimide reactions.

4.3 Characterization and Testing

4.3.1 Particle Size and Surface Charge

• 4.3.1.1 Measure the particle size using dynamic light scattering (DLS) and surface charge (zeta potential) to ensure the nanoparticles are within the desired range for targeting.

4.3.2 Drug Loading Efficiency

• 4.3.2.1 Quantify the amount of drug loaded into the nanoparticles using techniques such as UV-Vis spectrophotometry or high-performance liquid chromatography (HPLC).

4.3.3 In Vitro Targeting Efficacy

• 4.3.3.1 Perform in vitro studies on target cells (e.g., cancer cells) expressing the desired receptors to confirm the targeting capability of the nanoparticles.

4.4 Sterility and Storage

4.4.1 Sterilization

• 4.4.1.1 Sterilize the nanoparticle formulation by passing it through a 0.22 µm filter to remove contaminants.

4.4.2 Storage Conditions

• 4.4.2.1 Store the nanoparticles in sealed, sterile containers at 4°C to maintain stability and functionality of the targeting ligands.

5) Abbreviations, if any

• PLGA: Poly(lactic-co-glycolic acid)
• DLS: Dynamic Light Scattering
• EDC: 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide
• NHS: N-Hydroxysuccinimide

6) Documents, if any

• Nanoparticle Preparation Logbook
• Targeting Ligand Conjugation Logbook

7) References, if any

• Guidelines for nanoparticle preparation for drug delivery
• FDA Guidance on Nanotechnology

8) SOP Version

Version 1.0

Annexure

Nanoparticle Preparation Logbook Template

Date	Batch Number	Nanoparticle Material	Targeting Ligand	Particle Size	Drug Loading Efficiency	Operator Initials	QA Initials
DD/MM/YYYY	Batch Number	Material Name	Ligand Name	Size in nm	Efficiency (%)	Operator Name	QA Name

Targeting Ligand Conjugation Logbook Template

Date	Batch Number	Ligand Conjugated	Conjugation Method	Ligand Density	Binding Efficiency	Operator Initials	QA Initials
DD/MM/YYYY	Batch Number	Ligand Name	Method Used	Density (mol/mol)	Efficiency (%)	Operator Name	QA Name