Evidence-based standardized sample handling protocol for accurate blood-based Alzheimer's disease biomarker measurement : Results and consensus of the Global Biomarker Standardization Consortium
Verberk, Inge M.W. ; Gouda, Mariam ; Antwi-Berko, Daniel ; van Leeuwenstijn, Mardou ; Bongers, Bram ; Houtkamp, Isabel M. ; van der Flier, Wiesje M. ; Janelidze, Shorena LU ; Hansson, Oskar LU
and Bastard, Nathalie Le
, et al.
(2025)
In Alzheimer's and Dementia
21(10).
- Abstract
INTRODUCTION: Blood-based biomarkers (BBMs) have revolutionized Alzheimer's disease diagnosis and monitoring. Their pre-analytical stability requires scrutiny. This study assessed pre-analytical effects to inform a standardized sample handling protocol. METHODS: Assessed pre-analytical variations included collection tube type, hemolysis, centrifugation settings, centrifugation/storage delays, tube transfers, and freeze-thawing (n = 15/experiment). Phosphorylated tau (pTau) isoforms were measured with Simoa, Lumipulse, MesoScale Discovery, and immunoprecipitation-mass spectrometry. Amyloid-beta (Aβ42, Aβ40), glial fibrillary acidic protein (GFAP), and neurofilament light (NfL) protein were measured with Simoa. RESULTS: All assessed BBM... (More)
INTRODUCTION: Blood-based biomarkers (BBMs) have revolutionized Alzheimer's disease diagnosis and monitoring. Their pre-analytical stability requires scrutiny. This study assessed pre-analytical effects to inform a standardized sample handling protocol. METHODS: Assessed pre-analytical variations included collection tube type, hemolysis, centrifugation settings, centrifugation/storage delays, tube transfers, and freeze-thawing (n = 15/experiment). Phosphorylated tau (pTau) isoforms were measured with Simoa, Lumipulse, MesoScale Discovery, and immunoprecipitation-mass spectrometry. Amyloid-beta (Aβ42, Aβ40), glial fibrillary acidic protein (GFAP), and neurofilament light (NfL) protein were measured with Simoa. RESULTS: All assessed BBM levels varied by over 10% by collection tube type. Aβ peptides were the most sensitive, and their levels declined >by more than 10% under storage and centrifugation delays, more steeply at room temperature (RT) compared with 2°C to 8°C. NfL and GFAP levels increased by more than 10% upon RT/−20°C storage. pTau isoforms demonstrated stability across most pre-analytical variations. DISCUSSION: We established an evidence-based handling protocol to ensure reliable sample handling for neurological BBMs upon adoption in clinics, trials, and research. Highlights: Sample handling protocols can mitigate pre-analytical effects on BBM results. We developed an evidence-based, expert-consensus plasma sample handling protocol. Primary collection tube and delays to centrifuging or freezing impact AD BBMs. Plasma pTau217 is highly resistant to pre-analytical sample handling variations. Plasma Aβ42 and Aβ40 were most sensitive to pre-analytical variations.
(Less)
- author
- Verberk, Inge M.W.
; Gouda, Mariam
; Antwi-Berko, Daniel
; van Leeuwenstijn, Mardou
; Bongers, Bram
; Houtkamp, Isabel M.
; van der Flier, Wiesje M.
; Janelidze, Shorena
LU
; Hansson, Oskar
LU
and Bastard, Nathalie Le
, et al. (More)
- Verberk, Inge M.W.
; Gouda, Mariam
; Antwi-Berko, Daniel
; van Leeuwenstijn, Mardou
; Bongers, Bram
; Houtkamp, Isabel M.
; van der Flier, Wiesje M.
; Janelidze, Shorena
LU
; Hansson, Oskar
LU
; Bastard, Nathalie Le
; Vandijck, Manu
; Hunter, Jacob
; Honigberg, Lee
; Kirmess, Kristopher M.
; Verghese, Philip B.
; Blennow, Kaj
LU
; Zetterberg, Henrik
LU
; Meyers, Emily A.
; Edelmayer, Rebecca M.
and Teunissen, Charlotte
(Less)
- organization
- publishing date
- 2025-10
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- amyloid beta, glial fibrillary acidic protein, neurofilament light, phosphorylated tau, plasma biomarkers, pre-analytical variability, pre-analytics, sample handling, stability
- in
- Alzheimer's and Dementia
- volume
- 21
- issue
- 10
- article number
- e70752
- publisher
- Wiley
- external identifiers
-
- scopus:105017539165
- pmid:41025225
- ISSN
- 1552-5260
- DOI
- 10.1002/alz.70752
- language
- English
- LU publication?
- yes
- id
- 485e643d-1ca3-4e62-a632-04313b7200e4
- date added to LUP
- 2025-11-26 10:59:15
- date last changed
- 2025-12-10 20:12:46
@article{485e643d-1ca3-4e62-a632-04313b7200e4,
abstract = {{<p>INTRODUCTION: Blood-based biomarkers (BBMs) have revolutionized Alzheimer's disease diagnosis and monitoring. Their pre-analytical stability requires scrutiny. This study assessed pre-analytical effects to inform a standardized sample handling protocol. METHODS: Assessed pre-analytical variations included collection tube type, hemolysis, centrifugation settings, centrifugation/storage delays, tube transfers, and freeze-thawing (n = 15/experiment). Phosphorylated tau (pTau) isoforms were measured with Simoa, Lumipulse, MesoScale Discovery, and immunoprecipitation-mass spectrometry. Amyloid-beta (Aβ42, Aβ40), glial fibrillary acidic protein (GFAP), and neurofilament light (NfL) protein were measured with Simoa. RESULTS: All assessed BBM levels varied by over 10% by collection tube type. Aβ peptides were the most sensitive, and their levels declined >by more than 10% under storage and centrifugation delays, more steeply at room temperature (RT) compared with 2°C to 8°C. NfL and GFAP levels increased by more than 10% upon RT/−20°C storage. pTau isoforms demonstrated stability across most pre-analytical variations. DISCUSSION: We established an evidence-based handling protocol to ensure reliable sample handling for neurological BBMs upon adoption in clinics, trials, and research. Highlights: Sample handling protocols can mitigate pre-analytical effects on BBM results. We developed an evidence-based, expert-consensus plasma sample handling protocol. Primary collection tube and delays to centrifuging or freezing impact AD BBMs. Plasma pTau217 is highly resistant to pre-analytical sample handling variations. Plasma Aβ42 and Aβ40 were most sensitive to pre-analytical variations.</p>}},
author = {{Verberk, Inge M.W. and Gouda, Mariam and Antwi-Berko, Daniel and van Leeuwenstijn, Mardou and Bongers, Bram and Houtkamp, Isabel M. and van der Flier, Wiesje M. and Janelidze, Shorena and Hansson, Oskar and Bastard, Nathalie Le and Vandijck, Manu and Hunter, Jacob and Honigberg, Lee and Kirmess, Kristopher M. and Verghese, Philip B. and Blennow, Kaj and Zetterberg, Henrik and Meyers, Emily A. and Edelmayer, Rebecca M. and Teunissen, Charlotte}},
issn = {{1552-5260}},
keywords = {{amyloid beta; glial fibrillary acidic protein; neurofilament light; phosphorylated tau; plasma biomarkers; pre-analytical variability; pre-analytics; sample handling; stability}},
language = {{eng}},
number = {{10}},
publisher = {{Wiley}},
series = {{Alzheimer's and Dementia}},
title = {{Evidence-based standardized sample handling protocol for accurate blood-based Alzheimer's disease biomarker measurement : Results and consensus of the Global Biomarker Standardization Consortium}},
url = {{http://dx.doi.org/10.1002/alz.70752}},
doi = {{10.1002/alz.70752}},
volume = {{21}},
year = {{2025}},
}