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 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.

Citation

Georgios Katsikis, Iris E Hwang, Wade Wang, Vikas S Bhat, Nicole L McIntosh, Omair A Karim, Bartlomiej J Blus, Sha Sha, Vincent Agache, Jacqueline M Wolfrum, Stacy L Springs, Anthony J Sinskey, Paul W Barone, Richard D Braatz, Scott R Manalis. Weighing the DNA Content of Adeno-Associated Virus Vectors with Zeptogram Precision Using Nanomechanical Resonators. Nano letters. 2022 Feb 23;22(4):1511-1517

Mesh Tags

Substances

PMID: 35148107

search → result