Evidence map›Paper›PMID 35430965›Full record

ArticleProtein and peptide letters2022

Sample Processing Considerations for Protein Stability Studies of Low Concentration Biofluid Samples using Differential Scanning Calorimetry.

Gabriela Schneider, Nichola C Garbett

Abstract read
In one paragraph

Article in Protein and peptide letters, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

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1citing papers in PubMed
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1 · What the graph read from it

What it found

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2 · The registry

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3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Article
4 · The record

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5 · Who and what money

Authors and funding

2 authors.

Gabriela SchneiderUofL Health - Brown Cancer Center and Division of Medical Oncology and Hematology, Department of Medicine, University of Louisville, Louisville, KY 40202, USA.
Nichola C GarbettUofL Health - Brown Cancer Center and Division of Medical Oncology and Hematology, Department of Medicine, University of Louisville, Louisville, KY 40202, USA.

Funding

Enhanced diagnostic assessment in lupus using differential scanning calorimetryR01AI129959 · NIAID · UNIVERSITY OF LOUISVILLE · PI GARBETT, NICHOLA C. · 2017 to 2021
$1.8M
NIAID NIH HHS R01 AI129959
6 · The paper itself

Abstract

backgroundThe analysis of biofluid samples with low protein content (e.g., urine or saliva) can be challenging for downstream analysis methods with limited sensitivity. To circumvent this problem, sample processing methods are employed to increase the protein concentration in analyzed samples. However, for some techniques, like differential scanning calorimetry (DSC) that characterizes thermally-induced unfolding of biomolecules, sample processing must not affect native protein structure and stability.

methodsWe evaluated centrifugal concentration and stirred cell ultrafiltration, two common methods of sample concentration characterized by a low risk of protein denaturation, with the goal of establishing a protocol for DSC analysis of low concentration biospecimens.

resultsOur studies indicate that both methods can affect protein stability assessed by DSC and, even after optimization of several parameters, the obtained DSC profile (thermogram) suggested that sample processing affects the structure or intermolecular interactions of component proteins contributing to altered thermal stability detectable by DSC. We also found a relationship between changes in thermograms and low protein concentration, indicating that diluting biospecimens to concentrations below 0.1 mg/mL can perturb the intermolecular environment and affect the structure of proteins present in the solution.

conclusionDilution of samples below 0.1 mg/mL, as well as concentration of samples with low protein content, resulted in affected thermogram shapes suggesting changes in protein stability. This should be taken into account when concentrating dilute samples or employing techniques that lower the protein concentration (e.g., fractionation), when downstream applications include techniques, such as DSC, that require the preservation of native protein forms.

Indexed as

ProteinsSpecimen HandlingCalorimetry, Differential ScanningProtein DenaturationProtein StabilityProteinscentrifugal concentratorsdifferential scanning calorimetrylow protein biofluid samplesnative proteinSample concentrationsample processingstability studiesstirred cell ultrafiltration

Identifiers

PMID35430965
PMCPMC10353316

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