Improving Reproducibility to Meet Minimal Information for Studies of Extracellular Vesicles 2018 Guidelines in Nanoparticle Tracking Analysis
November 17th, 2021
Nanoparticle tracking analysis (NTA) is a widely used method to characterize extracellular vesicles. This paper highlights NTA experimental parameters and controls plus a uniform method of analysis and characterization of samples and diluents necessary to supplement the guidelines proposed by MISEV2018 and EV-TRACK for reproducibility between laboratories.
Overview
This article presents a protocol for nanoparticle tracking analysis (NTA) aimed at characterizing extracellular vesicles. It provides step-by-step instructions to ensure accurate and reproducible results, regardless of the operator's skill level.
Key Study Components
Area of Science
- Neuroscience
- Biology
- Extracellular Vesicle Research
Background
- NTA is a widely used method for characterizing extracellular vesicles.
- Quantification of nanoparticles remains a challenge in the field.
- Consistent results are crucial for reproducibility across laboratories.
- This protocol addresses the need for uniform analysis methods.
Purpose of Study
- To provide a reliable protocol for novice operators of the NanoSight LM10.
- To enhance reproducibility in extracellular vesicle research.
- To supplement existing guidelines for nanoparticle characterization.
Methods Used
- Step-by-step instructions for using the NanoSight LM10.
- Cleaning procedures for the laser module.
- Guidelines for sample preparation and analysis.
- Controls and experimental parameters for accurate results.
Main Results
- Consistent size and concentration analysis of nanoparticles.
- Improved accuracy in quantifying extracellular vesicles.
- Reproducible results across multiple trial runs.
- Enhanced understanding of NTA methodology.
Conclusions
- The protocol allows for reliable nanoparticle characterization.
- It addresses challenges in the extracellular vesicle research field.
- Following the protocol ensures consistent results regardless of user skill level.
Frequently Asked Questions
Executive Industry Relevance
Reproducible nanoparticle tracking analysis (NTA) is critical for robust extracellular vesicle (EV) characterization, directly impacting target validation and assay development in biopharma R&D. Adhering to MISEV2018 guidelines and transparent reporting standards reduces experimental ambiguity and supports confident decision-making at key discovery inflection points. Standardized NTA protocols enable reliable cross-study comparisons, strengthening portfolio-wide data integrity.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Supports rigorous hypothesis testing by enabling consistent EV quantification and sizing.
- Reduces biological variability, enhancing confidence in functional target validation.
- Facilitates mechanistic de-risking by controlling for sample and instrument variables.
Screening & Assay Development
- Prepares validated EV samples for downstream screening workflows.
- Enables assay standardization through detailed reporting of controls and instrument settings.
- Delivers quantitative outputs for reliable compound evaluation and platform reuse.
Translational & Preclinical Research
- Aligns EV characterization with translational biomarker requirements when reporting is comprehensive.
- Ensures continuity from discovery to preclinical validation by documenting critical experimental parameters.
- Supports risk-adjusted advancement decisions by minimizing uncontrolled technical variation.
Pipeline & Workflow Integration
NTA, when implemented with full reporting and controls, bridges early discovery and preclinical workflows by providing reproducible, quantitative EV data.
- Discovery Biology: Enables robust hypothesis testing and pathway clarification through standardized EV measurement.
- Screening: Provides reproducible, quantitative particle size and concentration data for assay readiness.
- Analytics: Generates statistical outputs that facilitate cross-condition and cross-study comparisons.
- Translational Research: Supports biomarker alignment and preclinical continuity when reporting meets MISEV2018 standards.
- Enterprise Reuse: Establishes a reusable protocol framework for consistent EV characterization across programs.
Operational & Enterprise Impact
- Scientific Value: Increases predictive confidence and reduces mechanistic ambiguity in EV studies.
- Operational Value: Promotes standardization, reproducibility, and scalability of NTA workflows.
- Strategic Value: Enables better go/no-go decisions and reduces late-stage biological risk through robust data.
- Portfolio Impact: Supports risk-adjusted prioritization and advancement of EV-related assets.
Implementation Considerations
- Requires operator training in NTA instrumentation and protocol adherence.
- Needs access to calibrated NTA systems and validated analytical software.
- Demands rigorous documentation of controls, instrument settings, and environmental variables.
- Must adapt protocols for different sample types and diluents as specified by guidelines.
- Limited by the need for comprehensive reporting to ensure reproducibility across sites.
Biopharma FAQs
Chapters in this video
0:04
Introduction
0:44
Cleaning and Assembly of the Laser Module
1:29
Flushing Procedure for the Laser Module Prior to and Between Samples
2:28
Focusing and Positioning of the Laser Module
3:34
Loading Standards/Samples/Diluent into the Laser Module for Nanoparticle Tracking Analysis (NTA)
4:28
Sample Nanoparticle Tracking Analysis (NTA)
6:06
Re-Analysis of the Current Sample at Different Detection Threshold
6:40
Results: Effects of Camera Level and Detection Threshold on Measured Particle Size and Concentration of Filtered and Unfiltered Samples
8:02
Conclusion
