TY - JOUR
T1 - Pre-analytical stability of serum biomarkers for neurological disease
T2 - Neurofilament-light, glial fibrillary acidic protein and contactin-1
AU - van Lierop, Zoë Y. G. J.
AU - Verberk, Inge M. W.
AU - van Uffelen, Kees W. J.
AU - Koel-Simmelink, Marleen J. A.
AU - in't Veld, Lisanne
AU - Killestein, Joep
AU - Teunissen, Charlotte E.
N1 - Funding Information: Competing interests: Z. Y. G. J. van Lierop, I. M. W. Verberk, C. W. J. van Uffelen, M. J. A. Koel-Simmelink and L. in ‘t Veld report no conflict of interest. J. Killestein has accepted speaker fees from Biogen, Roche, Teva, Sanofi, Merck, Genzyme, and Novartis. C. E. Teunissen has collaborations with ADx Neurosciences and Quanterix, and performed contract research for or received grants from the National MS Society, and from the Alzheimer’s Association, AxonNeurosciences, Biogen, Boehringer, Brainstorm Therapeutics, EIP Pharma, Esai, Janssen Prevention Center, Roche, Toyama and Vivoryon outside the submitted work. Research of C. E. Teunissen is supported by the European Commission (Marie Curie International Training Network, grant agreement No 860197 (MIRIADE), and JPND), Health Holland, the Dutch Research Council (ZonMW), Alzheimer Drug Discovery Foundation, the Selfridges Group Foundation, Alzheimer Netherlands, Alzheimer Association. Further, she is a recipient of ABOARD, which is a public-private partnership receiving funding from ZonMW (#73305095007) and Health Holland, Topsector Life Sciences & Health (PPP-allowance; #LSHM20106). More than 30 partners participate in ABOARD. ABOARD also receives funding from Edwin Bouw Fonds and Gieskes-Strijbisfonds. Also, C. E. Teunissen serves on the editorial board of Medidact Neurologie/Springer of the Neuromethods book series, on the editorial board of Alzheimer’s Research and Therapy and of Neurology; Neuroimmunology & Neuroinflammation. All funding is paid to her institution. Funding Information: Research funding: This project was funded by the National MS Society (Progressive MS Alliance Grant No. PA-2101-37214). The funding organization played no role in the study design; in the collection, analysis and interpretation of data; in the writing of the report; or in the decision to submit the report for publication. Funding Information: We thank all volunteers for donation of their blood for research purposes. We acknowledge Mr. H. (Harry) A. M. Twaalfhoven for his support and guidance during this project. The MS Center Amsterdam was supported by a program grant from the Dutch MS Research Foundation (18-358f). Publisher Copyright: © 2022 Zoë Y. G. J. van Lierop et al., published by De Gruyter, Berlin/Boston.
PY - 2022/5/1
Y1 - 2022/5/1
N2 - Objectives: Neurofilament-light (NfL), glial fibrillary acidic protein (GFAP) and contactin-1 (CNTN1) are blood-based biomarkers that could contribute to monitoring and prediction of disease and treatment outcomes in neurological diseases. Pre-analytical sample handling might affect results, which could be disease-dependent. We tested common handling variations in serum of volunteers as well as in a defined group of patients with multiple sclerosis (pwMS). Methods: Sample sets from 5 pwMS and 5 volunteers at the outpatient clinic were collected per experiment. We investigated the effect of the following variables: collection tube type, delayed centrifugation, centrifugation temperature, delayed storage after centrifugation and freeze-thawing. NfL and GFAP were measured by Simoa and CNTN1 by Luminex. A median recovery of 90-110% was considered stable. Results: For most pre-analytical variables, serum NfL and CNTN1 levels remained unaffected. In the total group, NfL levels increased (121%) after 6 h of delay at 2-8 °C until centrifugation, while no significant changes were observed after 24 h delay at room temperature (RT). In pwMS specifically, CNTN1 levels increased from additional freeze-thaw cycles number 2 to 4 (111%-141%), whereas volunteer levels remained stable. GFAP showed good stability for all pre-analytical variables. Conclusions: Overall, the serum biomarkers tested were relatively unaffected by variations in sample handling. For serum NfL, we recommend storage at RT before centrifugation at 2-8 °C up to 6 h or at RT up to 24 h. For serum CNTN1, we advise a maximum of two freeze-thaw cycles. Our results confirm and expand on recently launched consensus standardized operating procedures.
AB - Objectives: Neurofilament-light (NfL), glial fibrillary acidic protein (GFAP) and contactin-1 (CNTN1) are blood-based biomarkers that could contribute to monitoring and prediction of disease and treatment outcomes in neurological diseases. Pre-analytical sample handling might affect results, which could be disease-dependent. We tested common handling variations in serum of volunteers as well as in a defined group of patients with multiple sclerosis (pwMS). Methods: Sample sets from 5 pwMS and 5 volunteers at the outpatient clinic were collected per experiment. We investigated the effect of the following variables: collection tube type, delayed centrifugation, centrifugation temperature, delayed storage after centrifugation and freeze-thawing. NfL and GFAP were measured by Simoa and CNTN1 by Luminex. A median recovery of 90-110% was considered stable. Results: For most pre-analytical variables, serum NfL and CNTN1 levels remained unaffected. In the total group, NfL levels increased (121%) after 6 h of delay at 2-8 °C until centrifugation, while no significant changes were observed after 24 h delay at room temperature (RT). In pwMS specifically, CNTN1 levels increased from additional freeze-thaw cycles number 2 to 4 (111%-141%), whereas volunteer levels remained stable. GFAP showed good stability for all pre-analytical variables. Conclusions: Overall, the serum biomarkers tested were relatively unaffected by variations in sample handling. For serum NfL, we recommend storage at RT before centrifugation at 2-8 °C up to 6 h or at RT up to 24 h. For serum CNTN1, we advise a maximum of two freeze-thaw cycles. Our results confirm and expand on recently launched consensus standardized operating procedures.
KW - biomarkers
KW - immunoassay
KW - nervous system diseases
KW - pre-analytical phase
UR - http://www.scopus.com/inward/record.url?scp=85127919512&partnerID=8YFLogxK
U2 - https://doi.org/10.1515/cclm-2022-0007
DO - https://doi.org/10.1515/cclm-2022-0007
M3 - Article
C2 - 35333481
SN - 1434-6621
VL - 60
SP - 842
EP - 850
JO - Clinical chemistry and laboratory medicine
JF - Clinical chemistry and laboratory medicine
IS - 6
ER -