Marleen J A Koel-Simmelink1, Anke Vennegoor2, Joep Killestein3, Marinus A Blankenstein4, Niklas Norgren5, Carsten Korth6, Charlotte E Teunissen7. 1. Department of Clinical Chemistry, VU University Medical Center Amsterdam, PO Box 7057, 1007MB Amsterdam, The Netherlands. Electronic address: mja.koel-simmelink@vumc.nl. 2. Department of Neurology, VU University Medical Center Amsterdam, PO Box 7057, 1007MB Amsterdam. The Netherlands. Electronic address: a.vennegoor@vumc.nl. 3. Department of Neurology, VU University Medical Center Amsterdam, PO Box 7057, 1007MB Amsterdam. The Netherlands. Electronic address: j.killestein@vumc.nl. 4. Department of Clinical Chemistry, VU University Medical Center Amsterdam, PO Box 7057, 1007MB Amsterdam, The Netherlands. Electronic address: ma.blankenstein@vumc.nl. 5. UmanDiagnostics, PO Box 7996, 90719 Umeå, Sweden. Electronic address: niklas.norgren@umandiagnostics.se. 6. Neurodegeneration Unit, Department Neuropathology, University of Düsseldorf Medical School, Moorenstrase 5, 40225 Düsseldorf, Germany. Electronic address: ckorth@uni-duesseldorf.de. 7. Department of Clinical Chemistry, VU University Medical Center Amsterdam, PO Box 7057, 1007MB Amsterdam, The Netherlands. Electronic address: c.teunissen@vumc.nl.
Abstract
BACKGROUND: Neurofilament (Nf) proteins have been shown to be promising biomarkers for monitoring and predicting disease progression for various neurological diseases. The aim of this study was to evaluate the effects of pre-analytical variables on the concentration of neurofilament heavy (NfH) and neurofilament light (NfL) proteins. METHODS: For NfH an in-house newly-developed and validated SinglePlex Luminex assay was used; ELISA was used to analyze NfL. RESULTS: For the NfL ELISA assay, the intra- and inter-assay variation was respectively, 1.5% and 16.7%. Analytical performance of the NfH SinglePlex Luminex assay in terms of sensitivity (6.6pg/mL), recovery in cerebrospinal fluid (CSF) (between 90 and 104%), linearity (from 6.6-1250pg/mL), and inter- and intra-assay variation (<8%) were good. Concentrations of both NfL and NfH appeared not negatively affected by blood contamination, repeated freeze-thaw cycles (up to 4), delayed processing (up to 24hours) and during long-term storage at -20°C, 4°C, and room temperature. A decrease in concentration was observed during storage of both neurofilament proteins up to 21days at 37°C, which was significant by day 5. CONCLUSIONS: The newly developed NfH SinglePlex Luminex assay has a good sensitivity and is robust. Moreover, both NfH and NfL are stable under the most prevalent pre-analytical variations.
BACKGROUND: Neurofilament (Nf) proteins have been shown to be promising biomarkers for monitoring and predicting disease progression for various neurological diseases. The aim of this study was to evaluate the effects of pre-analytical variables on the concentration of neurofilament heavy (NfH) and neurofilament light (NfL) proteins. METHODS: For NfH an in-house newly-developed and validated SinglePlex Luminex assay was used; ELISA was used to analyze NfL. RESULTS: For the NfL ELISA assay, the intra- and inter-assay variation was respectively, 1.5% and 16.7%. Analytical performance of the NfH SinglePlex Luminex assay in terms of sensitivity (6.6pg/mL), recovery in cerebrospinal fluid (CSF) (between 90 and 104%), linearity (from 6.6-1250pg/mL), and inter- and intra-assay variation (<8%) were good. Concentrations of both NfL and NfH appeared not negatively affected by blood contamination, repeated freeze-thaw cycles (up to 4), delayed processing (up to 24hours) and during long-term storage at -20°C, 4°C, and room temperature. A decrease in concentration was observed during storage of both neurofilament proteins up to 21days at 37°C, which was significant by day 5. CONCLUSIONS: The newly developed NfH SinglePlex Luminex assay has a good sensitivity and is robust. Moreover, both NfH and NfL are stable under the most prevalent pre-analytical variations.
Authors: Alessandro Trentini; Manuel Comabella; Mar Tintoré; Marleen J A Koel-Simmelink; Joep Killestein; Birthe Roos; Alex Rovira; Carsten Korth; Philipp Ottis; Marinus A Blankenstein; Xavier Montalban; Tiziana Bellini; Charlotte E Teunissen Journal: J Neurol Date: 2014-09-17 Impact factor: 4.849
Authors: Marta Del Campo; Wesley Jongbloed; Harry A M Twaalfhoven; Robert Veerhuis; Marinus A Blankenstein; Charlotte E Teunissen Journal: Front Neurol Date: 2015-09-29 Impact factor: 4.003
Authors: Yolande A L Pijnenburg; Nicolaas A Verwey; Wiesje M van der Flier; Philip Scheltens; Charlotte E Teunissen Journal: Alzheimers Dement (Amst) Date: 2015-12-14
Authors: Yvonne W Van Dalen; Charlotte Blokhuis; Sophie Cohen; Jacqueline A Ter Stege; Charlotte E Teunissen; Jens Kuhle; Neeltje A Kootstra; Henriette J Scherpbier; Taco W Kuijpers; Peter Reiss; Charles B L M Majoie; Matthan W A Caan; Dasja Pajkrt Journal: Medicine (Baltimore) Date: 2016-03 Impact factor: 1.889
Authors: Everard G B Vijverberg; Annemiek Dols; Welmoed A Krudop; Marta Del Campo Milan; Cora J Kerssens; Flora Gossink; Niels D Prins; Max L Stek; Philip Scheltens; Charlotte E Teunissen; Yolande A L Pijnenburg Journal: Alzheimers Dement (Amst) Date: 2017-03-02
Authors: Lieke H Meeter; Elise G Dopper; Lize C Jiskoot; Raquel Sanchez-Valle; Caroline Graff; Luisa Benussi; Roberta Ghidoni; Yolande A Pijnenburg; Barbara Borroni; Daniela Galimberti; Robert Jr Laforce; Mario Masellis; Rik Vandenberghe; Isabelle Le Ber; Markus Otto; Rick van Minkelen; Janne M Papma; Serge A Rombouts; Mircea Balasa; Linn Öijerstedt; Vesna Jelic; Katrina M Dick; David M Cash; Sophie R Harding; M Jorge Cardoso; Sebastien Ourselin; Martin N Rossor; Alessandro Padovani; Elio Scarpini; Chiara Fenoglio; Maria C Tartaglia; Foudil Lamari; Christian Barro; Jens Kuhle; Jonathan D Rohrer; Charlotte E Teunissen; John C van Swieten Journal: Ann Clin Transl Neurol Date: 2016-07-01 Impact factor: 4.511