Octet® AAVX Biosensors for Rapid and Direct Quantitation of AAV Capsids

Rapid Quantitation of AAV Capsids for Various Serotypes for Bioprocess Development and Quality Control in Gene Therapy

Adeno-Associated virus (AAV) has been a vector of choice in gene therapy as a gene delivery tool. Multiple AAV serotypes including both native (wild type) and recombinant are used in part due to their tissue specificity (trophism). In the production and manufacture bioprocess workflow of AAVs, measuring the concentration of the virus capsid (viral particle) is an important quality attribute. Current methods used to quantitate virus capsid concentration, such as ELISA, Droplet Digital PCR (ddPCR), Analytical Ultracentrifugation (AUC) to name a few are both time consuming and laborious.

Overview

Herein, in this application note we present the results of the evaluation of the Octet® AAVX Biosensor. The AAVX Biosensors offer a quantitation dynamic range of 8.5E8 to 1.0E13 vp/mL, high precision (CV<10%) and broad AAV serotype binding specificity allowing for the quantitation of 10 different serotypes tested. Further, the AAVX Biosensors were observed to be compatible in different samples matrices encountered in upstream and downstream process intermediates and requiring minimal sample preparation.

Together with the Octet® Bio-Layer Interferometry (BLI) systems, the Octet® AAVX Biosensor quantitation assay workflow enables the rapid, real time and high-throughput measurement of AAV concentration in samples across the AAV bioprocess workflow enabling quick process optimization, quality check and increased productivity.

Document Type: Application Note
Page Count: 12
Read Time: 20 minutes
Last Updated: December 1, 2022
Authors: Yuanyuan Zhang, PhD, Ahmad Kour, Frederik Meierrieks, Benjamin Moritz Graf, PhD, Ivan Krylov, PhD, Ling Zhang, PhD, David Apiyo, PhD, Bob Dass, PhD

This Application Note Covers

  1. Rapid, direct, and label-free quantitation of AAV capsids in crude and purified samples
  2. Specificity to various AAV serotypes, including AAV1 – AAV9 and AAVrh10
  3. Direct quantitation of AAV capsids in the dynamic range of 8.5E8 – 1.0E13 vp/mL
  4. Efficient and cost-effective regeneration for biosensor reuse up to 20 times
  5. Good correlation of the results with ELISA

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