Weighing the DNA Content of Adeno-Associated Virus Vectors with Zeptogram Precision Using Nanomechanical Resonators
| Autor: | Anthony J. Sinskey, Scott R. Manalis, Vincent Agache, Iris E. Hwang, Jacqueline Wolfrum, Sha Sha, Richard D. Braatz, Wade Wang, Omair A. Karim, Nicole L. McIntosh, Paul W. Barone, Stacy L. Springs, Georgios Katsikis, Vikas Bhat, Bartlomiej J. Blus |
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| Rok vydání: | 2022 |
| Předmět: |
Materials science
Resolution (mass spectrometry) viruses Mechanical Engineering Genetic Vectors Bioengineering DNA General Chemistry Dependovirus medicine.disease_cause Condensed Matter Physics Virus Viral vector Analytical Ultracentrifugation chemistry.chemical_compound Capsid chemistry medicine Biomanufacturing General Materials Science Adeno-associated virus Biomedical engineering |
| Zdroj: | Nano Letters. 22:1511-1517 |
| ISSN: | 1530-6992 1530-6984 |
| DOI: | 10.1021/acs.nanolett.1c04092 |
| Popis: | Quantifying the composition of viral vectors used in vaccine development and gene therapy is critical for assessing their functionality. Adeno-Associated Virus (AAV) vectors, which are the most widely used viral vectors for in-vivo gene therapy, are typically characterized using PCR, ELISA, and Analytical Ultracentrifugation which require laborious protocols or hours of turnaround time. Emerging methods such as Charge-Detection Mass Spectroscopy, Static Light Scattering, and Mass Photometry offer turnaround times of minutes for measuring AAV mass, but mostly require purified AAV-based reference materials for calibration. Here, we demonstrate a method for using Suspended Nanomechanical Resonators (SNR) to directly measure both AAV mass and aggregation from a few microliters of sample within minutes. We achieve a resolution near 10 zeptograms which corresponds to 1% of the genome holding capacity of the AAV capsid. Our results show the potential of our method for providing real-time quality control of viral vectors during biomanufacturing. |
| Databáze: | OpenAIRE |
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