Evidence map›Paper›PMID 40204056›Full record

ArticleAnalytical biochemistry2025

Integrative biophysical characterization of molecular interactions: A case study with the sfGFP-nanobody complex.

Aysha K Demeler, James Bosco, Matthew J Sydor, Michelle Nemetchek, Saeed Mortezazadeh, Liam Kerr, Sophia Bird, Roza Gabdullina, Cindee Yates-Hansen, Levi J McClelland and 3 more

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Analytical biochemistry, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing 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

1 citing paper in PubMed.

  1. 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

13 authors.

Aysha K DemelerDepartment of Chemistry and Biochemistry, University of Lethbridge, Lethbridge, AB, T1K 3M4, Canada.
James BoscoDepartment of Chemistry and Biochemistry, University of Montana, Missoula, MT, 59812, USA.
Matthew J SydorDepartment of Chemistry and Biochemistry, University of Montana, Missoula, MT, 59812, USA; Division of Biological Sciences, University of Montana, Missoula, MT, 59812, USA.
Michelle NemetchekDepartment of Chemistry and Biochemistry, University of Montana, Missoula, MT, 59812, USA; Division of Biological Sciences, University of Montana, Missoula, MT, 59812, USA.
Saeed MortezazadehDepartment of Chemistry and Biochemistry, University of Lethbridge, Lethbridge, AB, T1K 3M4, Canada.
Liam KerrDepartment of Chemistry and Biochemistry, University of Lethbridge, Lethbridge, AB, T1K 3M4, Canada.
Sophia BirdDepartment of Chemistry and Biochemistry, University of Lethbridge, Lethbridge, AB, T1K 3M4, Canada.
Roza GabdullinaDepartment of Chemistry and Biochemistry, University of Lethbridge, Lethbridge, AB, T1K 3M4, Canada.
Cindee Yates-HansenDivision of Biological Sciences, University of Montana, Missoula, MT, 59812, USA.
Levi J McClellandDepartment of Chemistry and Biochemistry, University of Montana, Missoula, MT, 59812, USA; Division of Biological Sciences, University of Montana, Missoula, MT, 59812, USA.
Ekaterina VoroninaDivision of Biological Sciences, University of Montana, Missoula, MT, 59812, USA.
Trushar R PatelDepartment of Chemistry and Biochemistry, University of Lethbridge, Lethbridge, AB, T1K 3M4, Canada.
Borries DemelerDepartment of Chemistry and Biochemistry, University of Lethbridge, Lethbridge, AB, T1K 3M4, Canada; Department of Chemistry and Biochemistry, University of Montana, Missoula, MT, 59812, USA. Electronic address: demeler@gmail.com.

Funding

Surveillance genome sequencing to detect SARS-CoV-2 virus variants in MontanaP30GM140963 · NIGMS · UNIVERSITY OF MONTANA · PI BOWLER, BRUCE E · 2021 to 2025
$6.9M
Development of an UltraScan Meta-Scheduler for HPC Job SubmissionR01GM120600 · NIGMS · UNIVERSITY OF TEXAS HLTH SCIENCE CENTER · PI Emre H. Brookes, BORRIES DEMELER · 2016 to 2026
$3.6M
P granules and control of germ cells developmentR01GM109053 · NIGMS · UNIVERSITY OF MONTANA · PI VORONINA, EKATERINA · 2014 to 2022
$2.5M
NIGMS NIH HHS P30 GM140963NIGMS NIH HHS R01 GM109053NIGMS NIH HHS R01 GM120600
6 · The paper itself

Abstract

This study compares several analytical biophysical methods for investigating protein-protein interactions (PPIs) in solution, using the interaction between superfolder green fluorescent protein (sfGFP) and its anti-sfGFP nanobody enhancer as a model system. Techniques evaluated include microscale thermophoresis, fluorescence correlation spectroscopy, analytical ultracentrifugation with multi-wavelength and fluorescence detection, isothermal titration calorimetry, and analytical size exclusion chromatography coupled to multi-angle static light scattering and dynamic light scattering. Each method was assessed for information content, dynamic range, precision, and complementarity. The results consistently indicate a single-digit nanomolar dissociation constant and 1:1 stoichiometry for the interaction. While each technique offers unique insights into binding affinity, thermodynamics, and stoichiometry of the interaction, the multi-method approach provides a more complete and reliable characterization of PPIs. The study demonstrates how combining multiple complementary techniques enhances the robustness of PPI analysis in solution-phase conditions.

Indexed as

Green Fluorescent ProteinsSingle-Domain AntibodiesCalorimetryChromatography, GelProtein BindingSpectrometry, FluorescenceThermodynamicsUltracentrifugationGreen Fluorescent ProteinsSingle-Domain Antibodies

Identifiers

PMID40204056
PMCPMC12131814

What OpenQuestion holds

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LicenceCC BY-NC-ND
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.