A practical guide to immunoassay method validation
Open accessAbstract
Biochemical markers have a central position in the diagnosis and management of patients in clinical medicine, and also in clinical research and drug development, also for brain disorders, such as Alzheimer's disease. The enzyme-linked immunosorbent assay (ELISA) is frequently used for measurement of low-abundance biomarkers. However, the quality of ELISA methods varies, which may introduce both systematic and random errors. This urges the need for more rigorous control of assay performance, regardless of its use in a research setting, in clinical routine, or drug development. The aim of a method validation is to present objective evidence that a method fulfills the requirements for its intended use. Although much has been published on which parameters to investigate in a method validation, less is available on a detailed level on how to perform the corresponding experiments. To remedy this, standard operating procedures (SOPs) with step-by-step instructions for a number of different validation parameters is included in the present work together with a validation report template, which allow for a well-ordered presentation of the results. Even though the SOPs were developed with the intended use for immunochemical methods and to be used for multicenter evaluations, most of them are generic and can be used for other technologies as well.
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The in-house immunoassays for VAMP2 and SNAP25 were validated for analytical performance following established guidelines before clinical application.
“All in-house immunoassays were validated and analytically qualified prior to clinical application.”
CSF NPTX2 concentrations were measured using a newly developed ELISA that was validated according to international consensus criteria.
“CSF NPTX2 concentrations were measured using a newly developed ELISA from ADx Neurosciences (ADx Neurosciences NV, Belgium) that was validated according to international consensus criteria [54] at the Neurochemistry Laboratory of the Amsterdam University Medical Center in the Netherlands”
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Cites
The design of optimum multifactorial experiments
PLACKETT, R. L., & BURMAN, J. P. (1946) · Biometrika · doi:10.1093/biomet/33.4.305 · Cited by 3,906 · see citing works
Limit of detection. A closer look at the IUPAC definition
Long, G. L., & Winefordner, J. D. (1983) · Analytical Chemistry · doi:10.1021/ac00258a001 · Cited by 1,900 · see citing works
Cerebrospinal fluid and plasma biomarkers in Alzheimer disease
Blennow, K., Hampel, H., Weiner, M., & Zetterberg, H. (2010) · Nature Reviews Neurology · doi:10.1038/nrneurol.2010.4 · Cited by 1,863 · see citing works
Self-citationA multi-rule Shewhart chart for quality control in clinical chemistry.
Westgard, J. O., Barry, P. L., Hunt, M. R., & Groth, T. (1981) · Clinical Chemistry · doi:10.1093/clinchem/27.3.493 · Cited by 832 · see citing works
Fit-for-Purpose Method Development and Validation for Successful Biomarker Measurement
Lee, J. W., Devanarayan, V., Barrett, Y. C., WEINER, R. S., Allinson, J. L., Fountain, S. T., Keller, S., Weinryb, I., Green, M., Duan, L., et al. (2006) · Pharmaceutical Research · doi:10.1007/s11095-005-9045-3 · Cited by 770 · see citing works
Desilva, B. S., Smith, W. C., WEINER, R. S., Kelley, M. M., Smolec, J., Lee, B., Khan, M., Tacey, R. L., Hill, H. M., & Celniker, A. C. (2003) · Pharmaceutical Research · doi:10.1023/b:pham.0000003390.51761.3d · Cited by 624 · see citing works
Cerebrospinal fluid biomarkers in trials for Alzheimer and Parkinson diseases
Lleó, A., Cavedo, E., Parnetti, L., Vanderstichele, H., Herukka, S., Andreasen, N., Ghidoni, R., Lewczuk, P., Jeromin, A., Winblad, B. G., et al. (2014) · Nature Reviews Neurology · doi:10.1038/nrneurol.2014.232 · Cited by 159 · see citing works
Self-citationDetermination of parallelism and nonparallelism in bioassay dilution curves
Plikaytis, B. D., Holder, P. F., Pais, L. B., Maslanka, S., Gheesling, L. L., & Carlone, G. M. (1994) · Journal of Clinical Microbiology · doi:10.1128/jcm.32.10.2441-2447.1994 · Cited by 155 · see citing works
Valentin, M., Ma, S., Zhao, A., Legay, F., & Avraméas, A. (2011) · Journal of Pharmaceutical and Biomedical Analysis · doi:10.1016/j.jpba.2011.03.033 · Cited by 147 · see citing works
Bioanalytical Method Validation
(2014) · CRC Press eBooks · doi:10.1201/b17582-27 · Cited by 88 · see citing works
Verification/validation of the performances of analytical method
Vassault, A. J., Lesne-Hulin, A., Chapuzet, E., Arnaud, J., & Giroud, C. (2010) · Annales de biologie clinique · doi:10.1684/abc.2011.0562 · Cited by 20 · see citing works
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Research works printed but not linked: 1
- [12]CLSI. Evaluation of precision performance of quantitative measurement. methods; approved guideline-second edition. In: Wilhelm DM, editor. CLSI Document EP05-A2. Wayne, PA: Clinical and Laboratory Standards Institute (2004). p. 1–39.
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Verberk, I. M. W., Slot, R. E., Verfaillie, S. C. J., Heijst, H., Prins, N. D., van Berckel, B. N. M., Scheltens, P., Teunissen, C. E., & van der Flier, W. M. (2018) · Annals of Neurology · doi:10.1002/ana.25334 · Cited by 315 · see citing works
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MacLean, E. L., Wilson, S. R., Martin, W. L., Davis, J. M., Nazarloo, H. P., & Carter, C. S. (2019) · Psychoneuroendocrinology · doi:10.1016/j.psyneuen.2019.05.018 · Cited by 183 · see citing works
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Tabak, B. A., Leng, G., Szeto, A., Parker, K. J., Verbalis, J. G., Ziegler, T. E., Lee, M. R., Neumann, I. D., & Mendez, A. J. (2023) · Molecular Psychiatry · doi:10.1038/s41380-022-01719-z · Cited by 170 · see citing works
Teunissen, C. E., Elias, N., Koel‐Simmelink, M. J. A., Durieux‐Lu, S., Malekzadeh, A., Pham, T. V., Piersma, S. R., Beccari, T., Meeter, L. H. H., Dopper, E. G. P., et al. (2016) · Alzheimer's & Dementia: Diagnosis, Assessment & Disease Monitoring · doi:10.1016/j.dadm.2015.12.004 · Cited by 103 · see citing works
Self-citationHampel, H., Shaw, L. M., Aisen, P., Chen, C., Lleó, A., Iwatsubo, T., Iwata, A., Yamada, M., Ikeuchi, T., Jia, J., et al. (2022) · Alzheimer's & Dementia · doi:10.1002/alz.12372 · Cited by 77 · see citing works
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Gan, H.-W., Leeson, C., Aitkenhead, H., & Dattani, M. (2023) · Comprehensive Psychoneuroendocrinology · doi:10.1016/j.cpnec.2023.100188 · Cited by 15 · see citing works
Di Cerbo, A., Quagliano, N., Napolitano, A., Pezzuto, F., Iannitti, T., & Di Cerbo, A. (2021) · Diagnostics · doi:10.3390/diagnostics11091590 · Cited by 14 · see citing works
Glutamate receptor 4 as a fluid biomarker for the diagnosis of psychiatric disorders
Gómez de San José, N., Goossens, J., Al Shweiki, M. R., Halbgebauer, S., Oeckl, P., Steinacker, P., Danzer, K. M., Graf, H., Schönfeldt-Lecuona, C., Belbin, O., et al. (2022) · Journal of Psychiatric Research · doi:10.1016/j.jpsychires.2022.10.010 · Cited by 9 · see citing works
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- Authors
- Andreasson, U., Perret-Liaudet, A., van Waalwijk van Doorn, L. J. C., Blennow, K., Chiasserini, D., Engelborghs, S., Fladby, T., Genc, S., Kruse, N., Kuiperij, H. B., Kulic, L., Lewczuk, P., Mollenhauer, B., Mroczko, B., Parnetti, L., Vanmechelen, E., Verbeek, M. M., Winblad, B., Zetterberg, H., Koel-Simmelink, M., & Teunissen, C. E.
- Affiliations
- University of Gothenburg (SE)
- University of Perugia (IT)
- University of Antwerp (BE)
- University of Oslo (NO)
- Universitätsmedizin Göttingen (DE)
- Radboud University Nijmegen (NL)
- University of Zurich (CH)
- Friedrich-Alexander-Universität Erlangen-Nürnberg (DE)
- Medical University of Białystok (PL)
- ADx NeuroSciences (BE)
- Karolinska Institutet (SE)
- UCL Queen Square Institute of Neurology (GB)
- Amsterdam UMC Location Vrije Universiteit Amsterdam (NL)
- ORCID
- Venue
- Frontiers in Neurology
- Publisher
- Frontiers Media SA
- Published
- 2015-08-19
- Volume
- 6
- Pages
- 179-179
- ISSN
- 1664-2295 (ISSN-L)
- PMC
- PMC4541289
- OpenAlex
- W1789015615
- Cited by
- 546
- References
- 14
- Citations by year
- 2026: 61 · 2025: 58 · 2024: 64 · 2023: 47 · 2022: 61 · 2021: 67 · 2020: 60 · 2019: 37 · 2018: 42 · 2017: 30 · 2016: 17 · 2015: 2
- FWCI
- 22.35
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by
- Locations
- Frontiers in Neurology (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Amsterdam UMC (VU Amsterdam) - Institutional Repository (publishedVersion)
- Pure Amsterdam UMC (acceptedVersion, other-oa)
- Zurich Open Repository and Archive (University of Zurich) (acceptedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- UCL Discovery (University College London) (submittedVersion)
- GoeScholar The Publication Server of the Georg-August-Universität Göttingen (Georg-August-Universität Göttingen) (submittedVersion, cc-by)
- GRO (submittedVersion, cc-by)
- Radboud Repository (Radboud University) (submittedVersion)
- VUBIR (Vrije Universiteit Brussel) (submittedVersion)
- Zurich Open Repository and Archive (University of Zurich) (submittedVersion, cc-by)
- Radboud Repository (Radboud University) (submittedVersion, other-oa)
- Data Archiving and Networked Services (DANS) (submittedVersion, other-oa)
- Data Archiving and Networked Services (DANS) (submittedVersion)
- Zurich Open Repository and Archive (University of Zurich)
- Funders
- National Research Centre
- EU Joint Programme – Neurodegenerative Disease Research
- ZonMw
- Bundesministerium für Bildung und Forschung
- Belgian Federal Science Policy Office
- Fonds Wetenschappelijk Onderzoek
- Ministero della Salute
- Vetenskapsrådet
- Türkiye Bilimsel ve Teknolojik Araştırma Kurumu
- Norges Forskningsråd
- Universiteit Antwerpen
- Vlaamse regering
- Fondation pour la Recherche sur Alzheimer
- Krajowy Naukowy Osrodek Wiodacy
- Topics
- Health Sciences › Medicine › Physiology › Clinical Laboratory Practices and Quality Control
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced Biosensing Techniques and Applications
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical Methods in Pharmaceuticals
- Keywords
- ELISA, standard operating procedures, biochemical markers, biomarkers, multicenter evaluation, Alzheimer's disease, immunochemical methods
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