Evidence map›Paper›PMID 40490977›Full record

ArticleBiotechnology journal2025

Tracking Particle-Encapsulated DNA Across the Anion-Exchange Chromatography Fractions of Recombinant Adeno-Associated Virus Using Droplet Digital PCR and High-Throughput Sequencing.

Noriko Hashiba, Yuzhe Yuan, Emi Itou-Kudo, Kyoko Masumi-Koizumi, Keisuke Yusa, Kazuhisa Uchida

Abstract read
In one paragraph

Article in Biotechnology journal, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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1 · What the graph read from it

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2 · The registry

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4 · The record

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5 · Who and what money

Authors and funding

6 authors.

Noriko HashibaGraduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan.
Yuzhe YuanGraduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan.
Emi Itou-KudoGraduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan.
Kyoko Masumi-KoizumiGraduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan.
Keisuke YusaGraduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan.
Kazuhisa UchidaGraduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan.ORCID https://orcid.org/0000-0001-7360-4356

Funding

Japan Agency for Medical Research and Development JP18ae0201001
6 · The paper itself

Abstract

introductionSafe and effective adeno-associated virus (AAV) vectors are essential for gene therapy. Particle heterogeneity, specifically particle DNA of varying types and sizes, significantly affects recombinant adeno-associated virus (rAAV) performance. Previous studies have identified particle-associated DNAs; however, the specific DNA composition of these mixed populations remains poorly understood. This study aimed to investigate the DNA composition of the isolated subpopulations of rAAV particles obtained through anion exchange (AEX) chromatography.

methodsRAAV2-ZsGreen1 particles were fractionated on an AEX column, resulting in 12 distinct fractions. We analyzed the DNA composition of these fractions using droplet digital PCR (ddPCR) and MiSeq to identify the incorporated DNA heterogeneity in them.

resultsOur findings illustrated a clear trend in which the DNA content increasing in fractions was associated with an increased rAAV genomic DNA ratio of total particle DNA. The particle DNA content increased significantly across fractions from Peak 1 to Peak 2, showing approximately 30,000- and 5000-fold increases for ZsGreen1 (rAAV genome) and amp

conclusionsWith the elucidated profile of particle DNAs, this study provides detailed information on particle heterogeneity, shedding light on empty and partial particle formation and impurity DNA incorporation.

Indexed as

DependovirusDNA, ViralHigh-Throughput Nucleotide SequencingPolymerase Chain ReactionChromatography, Ion ExchangeGenetic VectorsHumansDNA, ViralAEX fractionsddPCRhigh‐throughput sequencingrAAV genomic and impurity DNArecombinant adeno‐associated virus

Identifiers

PMID40490977
PMCPMC12149490

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