Evidence map›Paper›PMID 39205208›Full record

ArticleViruses2024

In-Depth Comparison of Adeno-Associated Virus Containing Fractions after CsCl Ultracentrifugation Gradient Separation.

Mojca Janc, Kaja Zevnik, Ana Dolinar, Tjaša Jakomin, Maja Štalekar, Katarina Bačnik, Denis Kutnjak, Magda Tušek Žnidarič, Lorena Zentilin, Dmitrii Fedorov and 1 more

Abstract readComparative Study
In one paragraph

Article in Viruses, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.

0numbers the graph read from it
0cells of the map it votes in
3citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

3 citing papers in PubMed.

  1. Article
  2. Article
  3. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

11 authors.

Mojca JancNational Institute of Biology, Večna pot 121, 1000 Ljubljana, Slovenia.ORCID 0009-0003-1426-3274
Kaja ZevnikNational Institute of Biology, Večna pot 121, 1000 Ljubljana, Slovenia.
Ana DolinarNational Institute of Biology, Večna pot 121, 1000 Ljubljana, Slovenia.
Tjaša JakominNational Institute of Biology, Večna pot 121, 1000 Ljubljana, Slovenia.
Maja ŠtalekarNational Institute of Biology, Večna pot 121, 1000 Ljubljana, Slovenia.
Katarina BačnikNational Institute of Biology, Večna pot 121, 1000 Ljubljana, Slovenia.
Denis KutnjakNational Institute of Biology, Večna pot 121, 1000 Ljubljana, Slovenia.ORCID 0000-0002-5327-0587
Magda Tušek ŽnidaričNational Institute of Biology, Večna pot 121, 1000 Ljubljana, Slovenia.
Lorena ZentilinInternational Center for Genetic Engineering and Biotechnology, Area Science Park, Padriciano 99, 34149 Trieste, Italy.ORCID 0000-0001-7514-8155
Dmitrii FedorovDepartment of Bioproducts and Biosystems, Aalto University, P.O. Box 16100, 00076 Aalto, Finland.ORCID 0000-0002-3081-0878
David DobnikNational Institute of Biology, Večna pot 121, 1000 Ljubljana, Slovenia.ORCID 0000-0002-4698-6931

Funding

Slovenian Research Agency P4-0407; L4-3180; 1000-20-0105; 1000-21-0105; S-ZDR/22-27/0105
6 · The paper itself

Abstract

Recombinant adeno-associated viruses (rAAVs) play a pivotal role in the treatment of genetic diseases. However, current production and purification processes yield AAV-based preparations that often contain unwanted empty, partially filled or damaged viral particles and impurities, including residual host cell DNA and proteins, plasmid DNA, and viral aggregates. To precisely understand the composition of AAV preparations, we systematically compared four different single-stranded AAV (ssAAV) and self-complementary (scAAV) fractions extracted from the CsCl ultracentrifugation gradient using established methods (transduction efficiency, analytical ultracentrifugation (AUC), quantitative and digital droplet PCR (qPCR and ddPCR), transmission electron microscopy (TEM) and enzyme-linked immunosorbent assay (ELISA)) alongside newer techniques (multiplex ddPCR, multi-angle light-scattering coupled to size-exclusion chromatography (SEC-MALS), multi-angle dynamic light scattering (MADLS), and high-throughput sequencing (HTS)). Suboptimal particle separation within the fractions resulted in unexpectedly similar infectivity levels. No single technique could simultaneously provide comprehensive insights in the presence of both bioactive particles and contaminants. Notably, multiplex ddPCR revealed distinct vector genome fragmentation patterns, differing between ssAAV and scAAV. This highlights the urgent need for innovative analytical and production approaches to optimize AAV vector production and enhance therapeutic outcomes.

Indexed as

DependovirusUltracentrifugationVirionCentrifugation, Density GradientCesiumChloridesGenetic VectorsHEK293 CellsHumansTransduction, GeneticCesiumcesium chlorideChloridesanalytical methodsanalytical ultracentrifugation (AUC)CsCl ultracentrifugation gradientdigital droplet PCR (ddPCR)Illumina sequencingrecombinant adeno-associated viruses (rAAVs)size-exclusion chromatography coupled with multi-angle light scattering (SEC-MALS)transmission electron microscopy (TEM)

Identifiers

PMID39205208
PMCPMC11360810

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.