Evidence map›Paper›PMID 41282077›Full record

ArticleResearch square2025

Buffer Valency Engineering Enables High-concentration and Shelf-stable DNA Transfection Particles for Viral Vector Production.

Jinghan Lin, Yizong Hu, Turash H Pial, Kailei D Goodier, Di Yu, Marine Guise, Paetra Brailsford, Maria Choi-Ali, Sixuan Li, Yining Zhu and 7 more

Abstract readPreprint
PubMed Full text at PMC Publisher
In one paragraph

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

0numbers the graph read from it
0cells of the map it votes in
0citing 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

0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

5 · Who and what money

Authors and funding

17 authors.

Jinghan LinInstitute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA.ORCID 0009-0002-1475-1895
Yizong HuInstitute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA.ORCID 0000-0003-3983-5104
Turash H PialDepartment of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD, USA.
Kailei D GoodierInstitute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA.
Di YuInstitute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA.
Marine GuisePolyplus Sartorius, Illkirch-Graffenstaden, France.
Paetra BrailsfordBiogen Inc., Cambridge, MA, USA.
Maria Choi-AliBiogen Inc., Cambridge, MA, USA.
Sixuan LiInstitute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA.ORCID 0000-0002-0116-8812
Yining ZhuInstitute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA.ORCID 0000-0002-7178-0097
Jingyao MaInstitute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA.
Leonardo ChengInstitute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA.
Xiaoya LuInstitute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA.
Nicole KorinetzDepartment of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD, USA.
Tza-Huei WangInstitute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA.ORCID 0000-0002-3540-9354
Tine CurkDepartment of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD, USA.ORCID 0000-0002-2167-5336
Hai-Quan MaoInstitute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA.ORCID 0000-0002-4262-9988

Funding

Kinetic Assembly of Polymer-mRNA Nanoparticles Targets Circulating Monocytes to Enhance Cancer ImmunotherapyR01CA281143 · NCI · JOHNS HOPKINS UNIVERSITY · PI Jordan Green, Hai-Quan Mao · 2023 to 2026
$2.3M
NCI NIH HHS R01 CA281143
6 · The paper itself

Abstract

Cost-effective and scalable production is critical for advancing the clinical translation of adeno-associated virus (AAV)-mediated gene therapy. The widely used transient transfection method using plasmid DNA (pDNA)-loaded transfection particles for AAV production faces technical challenges due to instability of the particles and the concentration limits for particle preparation, hindering reproducibility and scalability. Here, we report a streamlined and scalable strategy to generate shelf-stable, highly concentrated pDNA/poly(ethylenimine) (PEI) transfection particles. By incorporating trivalent citrate ions in the dilution buffers, we kinetically modulate electrostatic complexation to achieve uniform nanoparticle assembly and prevent aggregation at high concentrations. This enables a tenfold increase in pDNA concentration in stabilized transfection particles from a typical range of 10-20 μg/mL to 200 μg/mL, while reducing the required dosing volume from 5-10% to 0.5% of the cell culture medium. The particle assembly process is robust to changes in mixing scale and timing and is compatible with standard workflows. We demonstrate equivalent AAV production efficiencies to standard methods and consistent performance in various production scales, which confirms the practical utility of this assembly method in developing robust, scalable, and cost-effective AAV manufacturing processes.

Indexed as

Adeno-associated virusKinetic controlled assemblyPlasmid DNAPoly(ethylenimine)Scalable manufacturingShelf-stable transfection particlesViral vector production

Identifiers

PMID41282077
PMCPMC12633500

What OpenQuestion holds

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LicenceCC BY
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Registered trials

None linked

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.