PMID- 34235314
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20210709
IS  - 2470-1343 (Electronic)
IS  - 2470-1343 (Linking)
VI  - 6
IP  - 25
DP  - 2021 Jun 29
TI  - Adeno-associated Virus Virus-like Particle Characterization via Orthogonal 
      Methods: Nanoelectrospray Differential Mobility Analysis, Asymmetric Flow 
      Field-Flow Fractionation, and Atomic Force Microscopy.
PG  - 16428-16437
LID - 10.1021/acsomega.1c01443 [doi]
AB  - Adeno-associated virus (AAV)-based virus-like particles (VLPs) are thriving 
      vectors of choice in the biopharmaceutical field of gene therapy. Here, a method 
      to investigate purified AAV serotype 8 (AAV8) batches via a nanoelectrospray 
      gas-phase mobility molecular analyzer (nES GEMMA), also known as an nES 
      differential mobility analyzer, is presented. Indeed, due to AAV's double-digit 
      nanometer scale, nES GEMMA is an excellently suited technique to determine the 
      surface-dry particle size termed electrophoretic mobility diameter of such VLPs 
      in their native state at atmospheric pressure and with particle-number-based 
      detection. Moreover, asymmetric flow field-flow fractionation (AF4, also known as 
      AFFFF) and atomic force microscopy (AFM) techniques were employed as orthogonal 
      techniques for VLP characterization. In addition, AF4 was implemented to 
      size-separate as well as to enrich and collect fractions of AAV8 VLPs after 
      inducing analyte aggregation in the liquid phase. Bionanoparticle aggregation was 
      achieved by a combination of heat and shear stress. These fractions were later 
      analyzed with nES GEMMA (in the gas phase) and AFM (on a solid surface). Both 
      techniques confirm the presence of dimers, trimers, and putative VLP oligomers. 
      Last, AFM reveals even larger AAV8 VLP aggregates, which were not detectable by 
      nES GEMMA because their heterogeneity combined with low abundance was below the 
      limit of detection of the instrument. Hence, the combination of the employed 
      orthogonal sizing methods with the separation technique AF4 allow a comprehensive 
      characterization of AAV8 VLPs applied as vectors.
CI  - (c) 2021 The Authors. Published by American Chemical Society.
FAU - Zoratto, Samuele
AU  - Zoratto S
AD  - Institute of Chemical Technologies and Analytics, TU Wien (Vienna University of 
      Technology), Vienna A-1060, Austria.
FAU - Weiss, Victor U
AU  - Weiss VU
AUID- ORCID: 0000-0002-0056-6819
AD  - Institute of Chemical Technologies and Analytics, TU Wien (Vienna University of 
      Technology), Vienna A-1060, Austria.
FAU - Friedbacher, Gernot
AU  - Friedbacher G
AD  - Institute of Chemical Technologies and Analytics, TU Wien (Vienna University of 
      Technology), Vienna A-1060, Austria.
FAU - Buengener, Carsten
AU  - Buengener C
AD  - Pharmaceutical Sciences, Baxalta Innovations (part of Takeda), Vienna A-1221, 
      Austria.
FAU - Pletzenauer, Robert
AU  - Pletzenauer R
AD  - Pharmaceutical Sciences, Baxalta Innovations (part of Takeda), Vienna A-1221, 
      Austria.
FAU - Foettinger-Vacha, Alexandra
AU  - Foettinger-Vacha A
AD  - Institute of Chemical Technologies and Analytics, TU Wien (Vienna University of 
      Technology), Vienna A-1060, Austria.
FAU - Graninger, Michael
AU  - Graninger M
AD  - Pharmaceutical Sciences, Baxalta Innovations (part of Takeda), Vienna A-1221, 
      Austria.
FAU - Allmaier, Guenter
AU  - Allmaier G
AUID- ORCID: 0000-0002-1438-9462
AD  - Institute of Chemical Technologies and Analytics, TU Wien (Vienna University of 
      Technology), Vienna A-1060, Austria.
LA  - eng
PT  - Journal Article
DEP - 20210615
PL  - United States
TA  - ACS Omega
JT  - ACS omega
JID - 101691658
PMC - PMC8246466
COIS- The authors declare the following competing financial interest(s): Carsten 
      Buengener, Robert Pletzenauer, Alexandra Foettinger-Vacha and Michael Graninger 
      are or were employees of Baxalta Innovations (part of Takeda).
EDAT- 2021/07/09 06:00
MHDA- 2021/07/09 06:01
PMCR- 2021/06/15
CRDT- 2021/07/08 06:46
PHST- 2021/03/17 00:00 [received]
PHST- 2021/05/27 00:00 [accepted]
PHST- 2021/07/08 06:46 [entrez]
PHST- 2021/07/09 06:00 [pubmed]
PHST- 2021/07/09 06:01 [medline]
PHST- 2021/06/15 00:00 [pmc-release]
AID - 10.1021/acsomega.1c01443 [doi]
PST - epublish
SO  - ACS Omega. 2021 Jun 15;6(25):16428-16437. doi: 10.1021/acsomega.1c01443. 
      eCollection 2021 Jun 29.