Glial fibrillary acidic protein in cerebrospinal fluid in humans is sensitive to various pre-analytical conditions: possible explanation and solution

Although glial fibrillary acidic protein (GFAP) has potential as a biomarker in cerebrospinal fluid, it is rarely used in clinical diagnosis due to high variability, low reliability, and poor reproducibility of results. Cerebrospinal fluid (CSF) was collected from patients (n = 167) at two sites at...

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Main Authors: Björn Evertsson, Ingela Nilsson Remahl, Max Albert Hietala, Anja Finn
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-08-01
Series:Frontiers in Neurology
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Online Access:https://www.frontiersin.org/articles/10.3389/fneur.2025.1627405/full
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author Björn Evertsson
Björn Evertsson
Ingela Nilsson Remahl
Max Albert Hietala
Max Albert Hietala
Anja Finn
author_facet Björn Evertsson
Björn Evertsson
Ingela Nilsson Remahl
Max Albert Hietala
Max Albert Hietala
Anja Finn
author_sort Björn Evertsson
collection DOAJ
description Although glial fibrillary acidic protein (GFAP) has potential as a biomarker in cerebrospinal fluid, it is rarely used in clinical diagnosis due to high variability, low reliability, and poor reproducibility of results. Cerebrospinal fluid (CSF) was collected from patients (n = 167) at two sites at the Department of Neurology. CSF was sampled in various volumes in both 10 mL polypropylene (PP) tubes and small, filled, sealed tubes of ≤2.0 mL (microtubes) for the comparison of GFAP concentrations. The influence of pH, sample volumes during storage and transport of CSF, under different temperatures, was tested to identify the losses and increase the possibilities of replicating data for GFAP. Concentrations of GFAP were measured by a sandwich ELISA. Exposure to air, agitation, and open-close cycles increased pH and lowered CO2. Compared to corresponding small filled sealed tubes, routine samples stored at −20°C showed 4–30% lower concentrations of GFAP. The loss increased further at lower volumes (< 0.5 mL). A significant difference in GFAP concentrations was seen in samples taken offsite (loss 42%) and onsite (loss 24%) compared to corresponding microtubes. Concentrations of GFAP remained stable in the microtubes, at 2–8°C and at RT for up to 3 weeks. GFAP in CSF is highly sensitive to changes in pH and dependent on adequate volumes for the best results. By avoiding exposure to air and agitation, we were able to stabilize GFAP concentrations in CSF by using small, filled, sealed tubes (microtubes). This handling could have impact on other biomarkers.
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spelling doaj-art-cc71fac0ee914bcb8d5972a073ec57a72025-08-20T03:58:11ZengFrontiers Media S.A.Frontiers in Neurology1664-22952025-08-011610.3389/fneur.2025.16274051627405Glial fibrillary acidic protein in cerebrospinal fluid in humans is sensitive to various pre-analytical conditions: possible explanation and solutionBjörn Evertsson0Björn Evertsson1Ingela Nilsson Remahl2Max Albert Hietala3Max Albert Hietala4Anja Finn5Department of Clinical Neuroscience, Karolinska Institute and Department of Neurology, Karolinska University Hospital, Stockholm, SwedenDepartment of Neurology, Laboratory for CSF Research, Karolinska University Hospital, Stockholm, SwedenDepartment of Clinical Neuroscience, Karolinska Institute and Department of Neurology, Karolinska University Hospital, Stockholm, SwedenDepartment of Clinical Neuroscience, Karolinska Institute and Department of Neurology, Karolinska University Hospital, Stockholm, SwedenDepartment of Neurology, Laboratory for CSF Research, Karolinska University Hospital, Stockholm, SwedenDepartment of Neurology, Laboratory for CSF Research, Karolinska University Hospital, Stockholm, SwedenAlthough glial fibrillary acidic protein (GFAP) has potential as a biomarker in cerebrospinal fluid, it is rarely used in clinical diagnosis due to high variability, low reliability, and poor reproducibility of results. Cerebrospinal fluid (CSF) was collected from patients (n = 167) at two sites at the Department of Neurology. CSF was sampled in various volumes in both 10 mL polypropylene (PP) tubes and small, filled, sealed tubes of ≤2.0 mL (microtubes) for the comparison of GFAP concentrations. The influence of pH, sample volumes during storage and transport of CSF, under different temperatures, was tested to identify the losses and increase the possibilities of replicating data for GFAP. Concentrations of GFAP were measured by a sandwich ELISA. Exposure to air, agitation, and open-close cycles increased pH and lowered CO2. Compared to corresponding small filled sealed tubes, routine samples stored at −20°C showed 4–30% lower concentrations of GFAP. The loss increased further at lower volumes (< 0.5 mL). A significant difference in GFAP concentrations was seen in samples taken offsite (loss 42%) and onsite (loss 24%) compared to corresponding microtubes. Concentrations of GFAP remained stable in the microtubes, at 2–8°C and at RT for up to 3 weeks. GFAP in CSF is highly sensitive to changes in pH and dependent on adequate volumes for the best results. By avoiding exposure to air and agitation, we were able to stabilize GFAP concentrations in CSF by using small, filled, sealed tubes (microtubes). This handling could have impact on other biomarkers.https://www.frontiersin.org/articles/10.3389/fneur.2025.1627405/fullCSFGFAPpre-analysismicrotubereliability
spellingShingle Björn Evertsson
Björn Evertsson
Ingela Nilsson Remahl
Max Albert Hietala
Max Albert Hietala
Anja Finn
Glial fibrillary acidic protein in cerebrospinal fluid in humans is sensitive to various pre-analytical conditions: possible explanation and solution
Frontiers in Neurology
CSF
GFAP
pre-analysis
microtube
reliability
title Glial fibrillary acidic protein in cerebrospinal fluid in humans is sensitive to various pre-analytical conditions: possible explanation and solution
title_full Glial fibrillary acidic protein in cerebrospinal fluid in humans is sensitive to various pre-analytical conditions: possible explanation and solution
title_fullStr Glial fibrillary acidic protein in cerebrospinal fluid in humans is sensitive to various pre-analytical conditions: possible explanation and solution
title_full_unstemmed Glial fibrillary acidic protein in cerebrospinal fluid in humans is sensitive to various pre-analytical conditions: possible explanation and solution
title_short Glial fibrillary acidic protein in cerebrospinal fluid in humans is sensitive to various pre-analytical conditions: possible explanation and solution
title_sort glial fibrillary acidic protein in cerebrospinal fluid in humans is sensitive to various pre analytical conditions possible explanation and solution
topic CSF
GFAP
pre-analysis
microtube
reliability
url https://www.frontiersin.org/articles/10.3389/fneur.2025.1627405/full
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