Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Halofantrine protects photoreceptors in multiple models of retinal degeneration.

Research square·2026
Same author

Dynamics of intronic polyadenylation in the hematopoietic lineage and its regulation by DNA methylation.

Genome research·2026
Same author

Sexual and Gender Minorities in Organ and Tissue Donation and Transplantation (OTDT): A Survey of Canadian OTDT Health Care Workers.

Canadian journal of kidney health and disease·2026
Same author

Start Making Sense: Addressing Reported Ambiguity of Final Margin Status for Oral Cavity Cancer.

Head & neck·2026
Same author

Intronic polyadenylation-derived long noncoding RNA modulates nucleolar integrity and function.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same author

Organ and tissue donation and transplantation in people who identify as Two-Spirit, gay, bisexual, transgender, queer, intersex, and more: a Canada-wide cross-sectional community survey.

Canadian journal of anaesthesia = Journal canadien d'anesthesie·2025

Related Experiment Video

Updated: May 21, 2025

Viability Assays for Cells in Culture
12:03

Viability Assays for Cells in Culture

Published on: January 20, 2014

46.1K

Validation of a quantitative cell-based relative potency assay for LUXTURNA.

Katherine A High1, Dave Le Blond2, Karen Doucette3

  • 1Spark Therapeutics, Philadelphia, PA 19103, USA.

Molecular Therapy. Methods & Clinical Development
|March 19, 2025
PubMed
Summary

A validated in vitro assay measures the biological activity of voretigene neparvovec-rzyl (Luxturna) by quantifying RPE65 protein production in HEK293 cells. This assay ensures lot-to-lot consistency for the first US-approved gene therapy for genetic eye disease.

Keywords:
AAV2-hRPE65v2LC-MS/MSLuxturnaassayin vitropotencyrelativevalidation

More Related Videos

High Throughput, Real-time, Dual-readout Testing of Intracellular Antimicrobial Activity and Eukaryotic Cell Cytotoxicity
09:09

High Throughput, Real-time, Dual-readout Testing of Intracellular Antimicrobial Activity and Eukaryotic Cell Cytotoxicity

Published on: November 16, 2016

7.8K
Author Spotlight: Developing Novel Anticancer Therapeutics Targeting the DNA Damage Response
05:01

Author Spotlight: Developing Novel Anticancer Therapeutics Targeting the DNA Damage Response

Published on: June 14, 2024

1.1K

Related Experiment Videos

Last Updated: May 21, 2025

Viability Assays for Cells in Culture
12:03

Viability Assays for Cells in Culture

Published on: January 20, 2014

46.1K
High Throughput, Real-time, Dual-readout Testing of Intracellular Antimicrobial Activity and Eukaryotic Cell Cytotoxicity
09:09

High Throughput, Real-time, Dual-readout Testing of Intracellular Antimicrobial Activity and Eukaryotic Cell Cytotoxicity

Published on: November 16, 2016

7.8K
Author Spotlight: Developing Novel Anticancer Therapeutics Targeting the DNA Damage Response
05:01

Author Spotlight: Developing Novel Anticancer Therapeutics Targeting the DNA Damage Response

Published on: June 14, 2024

1.1K

Area of Science:

  • * Gene Therapy
  • * Molecular Biology
  • * Assay Development

Background:

  • * Voretigene neparvovec-rzyl (Luxturna) is an AAV2 vector approved for treating inherited retinal diseases caused by RPE65 mutations.
  • * Regulatory approval necessitates validated assays demonstrating biological activity and potency.
  • * RPE65 protein is crucial for the visual cycle, converting all-trans-retinol to 11-cis-retinol.

Purpose of the Study:

  • * To validate an in vitro cell-based assay for measuring the biological activity and relative potency of voretigene neparvovec-rzyl (AAV2-hRPE65v2).
  • * To support regulatory approval and ensure lot-to-lot consistency of the gene therapy product.

Main Methods:

  • * Transduction of HEK293 cells with AAV2-hRPE65v2 vector.
  • * Quantification of vector-encoded RPE65 protein in cell lysates using liquid chromatography with tandem mass spectrometry (LC-MS/MS).
  • * Validation of the assay across seven key characteristics: suitability, specificity, linearity, precision, accuracy, range, and robustness.

Main Results:

  • * The validated assay accurately measures AAV2-hRPE65v2 transduction and subsequent RPE65 biological activity.
  • * The assay demonstrated reliability for assessing relative potency within a 50%-150% range of a reference standard.
  • * Successful validation supported lot-to-lot consistency, stability, and comparability assessments for Luxturna manufacturing.

Conclusions:

  • * This study reports the first validation of an in vitro cell-based relative potency assay for an AAV vector.
  • * The validated assay is critical for ensuring the quality and efficacy of Luxturna, the first gene therapy approved in the US for a genetic disease.
  • * The assay facilitates manufacturing process improvements and regulatory compliance for gene therapies.