Weighing the DNA Content of Adeno-Associated Virus Vectors with Zeptogram Precision Using Nanomechanical Resonators
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 anal...
| Main Authors: | , , , , , , , , , , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
| Published: |
American Chemical Society (ACS)
2023
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| Online Access: | https://hdl.handle.net/1721.1/146983 |
| _version_ | 1811084381411868672 |
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| author | Katsikis, Georgios Hwang, Iris E Wang, Wade Bhat, Vikas S McIntosh, Nicole L Karim, Omair A Blus, Bartlomiej J Sha, Sha Agache, Vincent Wolfrum, Jacqueline M Springs, Stacy L Sinskey, Anthony J Barone, Paul W Braatz, Richard D Manalis, Scott R |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Katsikis, Georgios Hwang, Iris E Wang, Wade Bhat, Vikas S McIntosh, Nicole L Karim, Omair A Blus, Bartlomiej J Sha, Sha Agache, Vincent Wolfrum, Jacqueline M Springs, Stacy L Sinskey, Anthony J Barone, Paul W Braatz, Richard D Manalis, Scott R |
| author_sort | Katsikis, Georgios |
| collection | MIT |
| description | 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 using optical or charge properties of AAV. Here, we demonstrate an orthogonal method where suspended nanomechanical resonators (SNR) are used to directly measure both AAV mass and aggregation from a few microliters of sample within minutes. We achieve a precision 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. |
| first_indexed | 2024-09-23T12:49:49Z |
| format | Article |
| id | mit-1721.1/146983 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T12:49:49Z |
| publishDate | 2023 |
| publisher | American Chemical Society (ACS) |
| record_format | dspace |
| spelling | mit-1721.1/1469832023-02-20T04:16:00Z Weighing the DNA Content of Adeno-Associated Virus Vectors with Zeptogram Precision Using Nanomechanical Resonators Katsikis, Georgios Hwang, Iris E Wang, Wade Bhat, Vikas S McIntosh, Nicole L Karim, Omair A Blus, Bartlomiej J Sha, Sha Agache, Vincent Wolfrum, Jacqueline M Springs, Stacy L Sinskey, Anthony J Barone, Paul W Braatz, Richard D Manalis, Scott R Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Mechanical Engineering Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Chemical Engineering 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 using optical or charge properties of AAV. Here, we demonstrate an orthogonal method where suspended nanomechanical resonators (SNR) are used to directly measure both AAV mass and aggregation from a few microliters of sample within minutes. We achieve a precision 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. 2023-01-05T18:47:24Z 2023-01-05T18:47:24Z 2022 2023-01-05T17:47:43Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/146983 Katsikis, Georgios, Hwang, Iris E, Wang, Wade, Bhat, Vikas S, McIntosh, Nicole L et al. 2022. "Weighing the DNA Content of Adeno-Associated Virus Vectors with Zeptogram Precision Using Nanomechanical Resonators." Nano Letters, 22 (4). en 10.1021/ACS.NANOLETT.1C04092 Nano Letters Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf American Chemical Society (ACS) MIT web domain |
| spellingShingle | Katsikis, Georgios Hwang, Iris E Wang, Wade Bhat, Vikas S McIntosh, Nicole L Karim, Omair A Blus, Bartlomiej J Sha, Sha Agache, Vincent Wolfrum, Jacqueline M Springs, Stacy L Sinskey, Anthony J Barone, Paul W Braatz, Richard D Manalis, Scott R Weighing the DNA Content of Adeno-Associated Virus Vectors with Zeptogram Precision Using Nanomechanical Resonators |
| title | Weighing the DNA Content of Adeno-Associated Virus Vectors with Zeptogram Precision Using Nanomechanical Resonators |
| title_full | Weighing the DNA Content of Adeno-Associated Virus Vectors with Zeptogram Precision Using Nanomechanical Resonators |
| title_fullStr | Weighing the DNA Content of Adeno-Associated Virus Vectors with Zeptogram Precision Using Nanomechanical Resonators |
| title_full_unstemmed | Weighing the DNA Content of Adeno-Associated Virus Vectors with Zeptogram Precision Using Nanomechanical Resonators |
| title_short | Weighing the DNA Content of Adeno-Associated Virus Vectors with Zeptogram Precision Using Nanomechanical Resonators |
| title_sort | weighing the dna content of adeno associated virus vectors with zeptogram precision using nanomechanical resonators |
| url | https://hdl.handle.net/1721.1/146983 |
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