Article
Chemistry, Medicinal
Takahiro Maruno, Kentaro Ishii, Tetsuo Torisu, Susumu Uchiyama
Summary: This study presents an effective approach for size distribution analysis and component identification of AAV vector samples using a small amount of material. The proposed band sedimentation analytical ultracentrifugation (BS-AUC) method offers higher resolution and allows for accurate determination of component concentration compared to traditional sedimentation velocity AUC (SV-AUC). This approach overcomes the sample amount limitation of SV-AUC.
JOURNAL OF PHARMACEUTICAL SCIENCES
(2023)
Article
Medicine, Research & Experimental
Harshit Khasa, Greg Kilby, Xiaoyu Chen, Chunlei Wang
Summary: ABC was evaluated for separation of therapeutic AAVs, showing effective separation between full and empty AAV particles. Results demonstrated consistent separation and quantification, with reproducibility and insensitivity to experimental variations. The technique shows promise for routine analysis in therapeutic products.
MOLECULAR THERAPY-METHODS & CLINICAL DEVELOPMENT
(2021)
Article
Biotechnology & Applied Microbiology
Amy Henrickson, Xiaozhe Ding, Austin G. Seal, Zhe Qu, Lauren Tomlinson, John Forsey, Viviana Gradinaru, Kazuhiro Oka, Borries Demeler
Summary: This study presents guidelines for experimental design of multi-wavelength analytical ultracentrifugation (MW-AUC) and compares its results with other methods. The results demonstrate that MW-AUC provides more accurate characterization and quantification of adeno-associated viruses, and identifies contaminants.
Review
Medicine, Research & Experimental
Andreas L. Gimpel, Georgios Katsikis, Sha Sha, Andrew John Maloney, Moo Sun Hong, Tam N. T. Nguyen, Jacqueline Wolfrum, Stacy L. Springs, Anthony J. Sinskey, Scott R. Manalis, Paul W. Barone, Richard D. Braatz
Summary: The article introduces analytical methods for rAAV quality attributes, emphasizing the importance of rapid, high-throughput methods, which are not yet widely used in academic and industrial practice, and have limited supporting data.
MOLECULAR THERAPY-METHODS & CLINICAL DEVELOPMENT
(2021)
Article
Biophysics
Sylvie Saleun, Caroline Mas, Aline Le Roy, Magalie Penaud-Budloo, Oumeya Adjali, Veronique Blouin, Christine Ebel
Summary: Recombinant adeno-associated virus virus-derived vectors (rAAVs) are widely used in vivo gene therapies due to their safety profile. However, rAAV production often results in a heterogeneous vector population with undesired empty particles. Analytical ultracentrifugation sedimentation velocity (AUC-SV) is a gold standard technique to measure the relative amounts of each vector subpopulation and components. This letter discusses the principle and practice of using AUC for rAAVs characterization, including the use of interference optics to estimate weight percentages of different capsid populations and the genome size incorporated in rAAV particles.
EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS
(2023)
Article
Chemistry, Medicinal
Takahiro Maruno, Kaede Usami, Kentaro Ishii, Tetsuo Torisu, Susumu Uchiyama
Summary: A multiwavelength sedimentation velocity analytical ultracentrifugation method was developed to characterize different types of particles and impurities in adeno-associated virus vector samples. This method can help reduce impurities in vector samples and establish a comprehensive approach for quantifying viral vector particles and related impurities.
JOURNAL OF PHARMACEUTICAL SCIENCES
(2021)
Article
Biophysics
Alexey Savelyev, Emre H. Brookes, Amy Henrickson, Borries Demeler
Summary: This study presents a method for characterizing and quantifying peaks formed in an analytical buoyant density equilibrium (ABDE) experiment. An algorithm is derived to calculate the concentration of the density forming gradient material at every point in the cell, provided specific parameters are known. Additionally, a new peak fitting algorithm has been developed to quantify the peaks in terms of density, apparent partial specific volume, and relative abundance.
EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS
(2023)
Article
Chemistry, Analytical
Durgadevi Perumal, Manikandan Krishnan, K. S. Lakshmi
Summary: Molnupiravir, a novel oral broad-spectrum antiviral drug, was quantified using UV spectrophotometric and high-performance liquid chromatographic methods. The suggested techniques meet the standards of ICH Q2 (R1) and have a wide linearity range and high accuracy.
ANALYTICAL CHEMISTRY LETTERS
(2023)
Article
Biochemistry & Molecular Biology
Shawn M. Sternisha, Peilu Liu, Alan G. Marshall, Brian G. Miller
Article
Chemistry, Multidisciplinary
Artur K. Mailyan, Joanna L. Chen, Weiwei Li, Arturo A. Keller, Shawn M. Sternisha, Brian G. Miller, Armen Zakarian
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
(2018)
Article
Materials Science, Biomaterials
Ross VerHeul, Craig Sweet, David H. Thompson
BIOMATERIALS SCIENCE
(2018)
Article
Biochemistry & Molecular Biology
Aditya Kulkarni, Vivek Badwaik, Kyle DeFrees, Ryan A. Schuldt, Dinara S. Gunasekera, Cory Powers, Alexander Vlahu, Ross VerHeul, David H. Thompson
Article
Cell Biology
Aditya Kulkarni, Kyle DeFrees, Ryan A. Schuldt, Alexander Vlahu, Ross VerHeul, Seok-Hee Hyun, Wei Deng, David H. Thompson
INTEGRATIVE BIOLOGY
(2013)
Article
Medicine, Research & Experimental
Aditya Kulkarni, Kyle DeFrees, Ryan A. Schuldt, Seok-Hee Hyun, Kyle J. Wright, Charu K. Yerneni, Ross VerHeul, David H. Thompson
MOLECULAR PHARMACEUTICS
(2013)
Review
Biochemistry & Molecular Biology
Shawn M. Sternisha, Brian G. Miller
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
(2019)
Article
Biophysics
Shawn M. Sternisha, A. Carl Whittington, Juliana A. Martinez Fiesco, Carol Porter, Malcolm M. McCray, Timothy Logan, Cristina Olivieri, Gianluigi Veglia, Peter J. Steinbach, Brian G. Miller
BIOPHYSICAL JOURNAL
(2020)
Article
Chemistry, Multidisciplinary
Rebecca Notis Dardashti, Shailesh Kumar, Shawn M. Sternisha, Post Sai Reddy, Brian G. Miller, Norman Metanis
CHEMISTRY-A EUROPEAN JOURNAL
(2020)
Article
Biochemical Research Methods
Shawn M. Sternisha, Prabuddha Mukherjee, Aneesh Alex, Eric J. Chaney, Ronit Barkalifa, Boyong Wan, Jang Hyuk Lee, Jose Rico-Jimenez, Mantas Zurauskas, Darold R. Spillman, Sobhana A. Sripada, Marina Marjanovic, Zane Arp, Sybille S. Galosy, Dharmesh S. Bhanushali, Steve R. Hood, Sayantan Bose, Stephen A. Boppart
Summary: This study used two-photon fluorescence lifetime imaging microscopy (2P-FLIM) to directly probe the health of CHO cells from a bioreactor, showing the potential of this technique for monitoring changes in the cellular microenvironment during cell culture processes. Analysis of FLIM data revealed differences in fluorescence lifetime parameters among different cell lines, serving as an indicator of metabolic activity changes.
BIOTECHNOLOGY JOURNAL
(2021)
Article
Biophysics
Shawn M. Sternisha, Abraham D. Wilson, Emilie Bouda, Akash Bhattacharya, Ross VerHeul
Summary: Accurate characterization of capsid species is crucial for viral vector-based gene therapies and vaccines. The current standard method, sedimentation velocity analytical ultracentrifugation (SV-AUC), is often size-limited and requires specialized software for analysis. The density gradient equilibrium AUC (DGE-AUC) provides a simplified and high-resolution method for separating biologics of different densities. It offers improvements in sensitivity and can be used with larger viral particles. This serotype-agnostic method enables intuitive interpretation and analysis without the need for specialized software.
EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS
(2023)
Meeting Abstract
Biochemistry & Molecular Biology
Akash Bhattacharya, Nicholas Harrington, Ross Verheul, Eric Von Seggern, Stephen Otts