A Practical Guide to Immunoassay Method Validation.
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.
Clinical Neurochemistry Laboratory, Institute of Neuroscience and Physiology, The Sahlgrenska Academy, University of Gothenburg , Mölndal , Sweden.
Ulf
U
Andreasson
Neurobiology Laboratory, Centre for Memory Resources and Research (CMRR), Groupement Hospitalier Est (GHE), Hôpitaux de Lyon, Université Lyon 1, CNRS UMR5292, INSERM U1028 , Lyon , France.
Armand
A
Perret-Liaudet
Department of Neurology, Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center , Nijmegen , Netherlands ; Department of Laboratory Medicine, Donders Institute for Brain, Cognition and Behaviour, Radboud Alzheimer Centre , Nijmegen , Netherlands.
Linda J C
LJ
van Waalwijk van Doorn
Clinical Neurochemistry Laboratory, Institute of Neuroscience and Physiology, The Sahlgrenska Academy, University of Gothenburg , Mölndal , Sweden.
Kaj
K
Blennow
Laboratory of Clinical Neurochemistry, Department of Medicine, Section of Neurology, University of Perugia , Perugia , Italy.
Davide
D
Chiasserini
Reference Center for Biological Markers of Dementia (BIODEM), Institute Born-Bunge, University of Antwerp , Antwerp , Belgium ; Department of Neurology and Memory Clinic, Hospital Network Antwerp (ZNA) Middelheim and Hoge Beuken , Antwerp , Belgium.
Sebastiaan
S
Engelborghs
University of Oslo , Oslo , Norway ; Department of Neurology, Akershus University Hospital , Lørenskog , Norway.
Tormod
T
Fladby
Department of Neuroscience, Institute of Health Science, Dokuz Eylul University , Izmir , Turkey.
Sermin
S
Genc
Department of Neuropathology, University Medical Center , Göttingen , Germany.
Niels
N
Kruse
Department of Neurology, Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center , Nijmegen , Netherlands ; Department of Laboratory Medicine, Donders Institute for Brain, Cognition and Behaviour, Radboud Alzheimer Centre , Nijmegen , Netherlands.
H Bea
HB
Kuiperij
Division of Psychiatry Research, University of Zurich , Schlieren , Switzerland.
Luka
L
Kulic
Department of Psychiatry and Psychotherapy, Universitätsklinikum Erlangen, Friedrich-Alexander Universität Erlangen-Nürnberg , Erlangen , Germany.
Piotr
P
Lewczuk
Paracelsus-Elena Klinik , Kassel , Germany ; Department of Neuropathology, University Medical Center Göttingen , Göttingen , Germany.
Brit
B
Mollenhauer
Department of Neurodegeneration Diagnostics, Medical University of Białystok , Białystok , Poland.
Barbara
B
Mroczko
Laboratory of Clinical Neurochemistry, Department of Medicine, Section of Neurology, University of Perugia , Perugia , Italy.
Lucilla
L
Parnetti
ADx NeuroSciences , Ghent , Belgium.
Eugeen
E
Vanmechelen
Department of Neurology, Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center , Nijmegen , Netherlands ; Department of Laboratory Medicine, Donders Institute for Brain, Cognition and Behaviour, Radboud Alzheimer Centre , Nijmegen , Netherlands.
Marcel M
MM
Verbeek
Karolinska Institutet , Stockholm , Sweden.
Bengt
B
Winblad
Clinical Neurochemistry Laboratory, Institute of Neuroscience and Physiology, The Sahlgrenska Academy, University of Gothenburg , Mölndal , Sweden ; Institute of Neurology, University College London , London , UK.
Henrik
H
Zetterberg
Neurochemistry Laboratory and Biobank, Department of Clinical Chemistry, Neurocampus Amsterdam, VU University Medical Center , Amsterdam , Netherlands.
Marleen
M
Koel-Simmelink
Neurochemistry Laboratory and Biobank, Department of Clinical Chemistry, Neurocampus Amsterdam, VU University Medical Center , Amsterdam , Netherlands.
Charlotte E
CE
Teunissen
immunoassays; limits of quantitation; method validation; precision; robustness