Evidence map›Paper›PMID 35157287›Full record

ArticleMethods in molecular biology (Clifton, N.J.)2022

Visualizing Filoviral Nucleoproteins Using Nanobodies Fused to the Ascorbate Peroxidase Derivatives APEX2 and dEAPX.

Laura Jo Sherwood, Andrew Hayhurst

Abstract read
In one paragraph

Article in Methods in molecular biology (Clifton, N.J.), 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.

0numbers the graph read from it
0cells of the map it votes in
5citing papers in PubMed
6.5field-weighted citation impact, top 3% of its field
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

5 citing papers in PubMed, 7 citations in OpenAlex.

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  5. Review
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

2 authors at 1 institution in 1 country.

Laura Jo SherwoodDisease Intervention and Prevention, Texas Biomedical Research Institute, San Antonio, TX, USA.
Andrew HayhurstDisease Intervention and Prevention, Texas Biomedical Research Institute, San Antonio, TX, USA. ahayhurst@TxBiomed.org.
Texas Biomedical Research Institute · US

Funding

Mechanism and Evolution of Filoviral Monoclonal Affinity Reagent Sandwich AssaysR01AI112851 · NIAID · TEXAS BIOMEDICAL RESEARCH INSTITUTE · PI HAYHURST, ANDREW · 2015 to 2019
$2.4M
NIAID NIH HHS R01 AI112851
6 · The paper itself

Abstract

Fusions of single-domain antibodies (sdAbs, nanobodies) to enzymatic reporters make convenient molecular probes to detect the presence of an antigen of interest. We have previously fused the monomeric hyperactive ascorbate peroxidase derivative APEX2 to anti-Ebolavirus and anti-Marburgvirus sdAbs to generate immunoreagents useful in detecting nucleoprotein (NP) on western blots, ELISA, and within cells following transfection of NP expression plasmids or following virus infection. Here we present the methods used to overexpress and purify these sdAb-APEX2 fusion proteins, and to employ them as probes in various scenarios with colorimetric and fluorometric signal development. We also introduce a dimeric hyperactive ascorbate peroxidase derivative dEAPX that enables bivalent sdAb probes to be produced while avoiding the need to generate more complex tandem sdAbs, leveraging avidity for improved signal strength. The APEX2 and dEAPX reagents appear interchangeable with any existing detection platform and the methods described here should enable a user to study their antigen of interest by simply swapping out the sdAb for their recombinant affinity reagent of choice.

Indexed as

EbolavirusMarburgvirusSingle-Domain AntibodiesAscorbate PeroxidasesNucleoproteinsAscorbate PeroxidasesNucleoproteinsSingle-Domain AntibodiesAPEX2Ascorbate peroxidaseFilovirusFocus forming assayMicroscopyNanobodyNucleoproteinsdAbVHHVirus-infected cellWestern blot

Identifiers

PMID35157287
PMCPMC13089185
OpenAlexW4213182647

What OpenQuestion holds

Textmetadata
LicenceTDM
Read underepoch 390

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.