Neurofilament ELISA validation

Axel Petzold, Ayse Altintas, Laura Andreoni, Ales Bartos, Achim Berthele, Marinus A. Blankenstein, Luc Buee, Massimiliano Castellazzi, Sabine Cepok, Manuel Comabella, Cris S. Constantinescu, Florian Deisenhammer, Gunnur Deniz, Gaye Erten, Mercedes Espiño, Enrico Fainardi, Diego Franciotta, Mark S. Freedman, Vilmantas Giedraitis, Nils Erik GilhusGavin Giovannoni, Andrzej Glabinski, Pawel Grieb, Hans Peter Hartung, Bernhard Hemmer, Sanna Kaisa Herukka, Rogier Hintzen, Martin Ingelsson, Samuel Jackson, Steve Jacobsen, Naghmeh Jafari, Marcin Jalosinski, Sven Jarius, Elisabeth Kapaki, Bernd C. Kieseier, Marleen J A Koel-Simmelink, Johannes Kornhuber, Jens Kuhle, Jacek Kurzepa, Patrice H. Lalive, Lars Lannfelt, Vera Lehmensiek, Piotr Lewczuk, Paolo Livrea, Fabiana Marnetto, Davide Martino, Til Menge, Niklas Norgren, Eva Papuć, George P. Paraskevas, Tuula Pirttilä, Cecília Rajda, Konrad Rejdak, Jan Ricny, Daniela Ripova, Lars Rosengren, Maddalena Ruggieri, Susanna Schraen, Gerry Shaw, Christian Sindic, Aksel Siva, Torgny Stigbrand, Iva Stonebridge, Baris Topcular, Maria Trojano, Hayrettin Tumani, Harry A M Twaalfhoven, L. Vécsei, Vincent Van Pesch, Hugo Vanderstichele, Christian Vedeler, Marcel M. Verbeek, Luisa Maria Villar, Robert Weissert, Brigitte Wildemann, Cui Yang, Karen Yao, Charlotte E. Teunissen

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Background: Neurofilament proteins (Nf) are highly specific biomarkers for neuronal death and axonal degeneration. As these markers become more widely used, an inter-laboratory validation study is required to identify assay criteria for high quality performance. Methods: The UmanDiagnostics NF-light ®enzyme-linked immunoabsorbent assays (ELISA) for the neurofilament light chain (NfL, 68 kDa) was used to test the intra-assay and inter-laboratory coefficient of variation (CV) between 35 laboratories worldwide on 15 cerebrospinal fluid (CSF) samples. Critical factors, such as sample transport and storage, analytical delays, reaction temperature and time, the laboratories' accuracy and preparation of standards were documented and used for the statistical analyses. Results: The intra-laboratory CV averaged 3.3% and the inter-laboratory CV 59%. The results from the test laboratories correlated with those from the reference laboratory (R = 0.60, p <0.0001). Correcting for critical factors improved the strength of the correlation. Differences in the accuracy of standard preparation were identified as the most critical factor. Correcting for the error introduced by variation in the protein standards improved the correlation to R = 0.98, p <0.0001 with an averaged inter-laboratory CV of 14%. The corrected overall inter-rater agreement was subtantial (0.6) according to Fleiss' multi-rater kappa and Gwet's AC1 statistics. Conclusion: This multi-center validation study identified the lack of preparation of accurate and consistent protein standards as the main reason for a poor inter-laboratory CV. This issue is also relevant to other protein biomarkers based on this type of assay and will need to be solved in order to achieve an acceptable level of analytical accuracy. The raw data of this study is available online.

Original languageEnglish
Pages (from-to)23-31
Number of pages9
JournalJournal of Immunological Methods
Volume352
Issue number1-2
DOIs
Publication statusPublished - Jan 31 2010

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Intermediate Filaments
Enzymes
Validation Studies
Biomarkers
Neurofilament Proteins
Light
Proteins
Cerebrospinal Fluid
Temperature

Keywords

  • Biomarker
  • NEFL
  • Neurofilament
  • NF-L
  • NfL
  • Validation

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

Cite this

Petzold, A., Altintas, A., Andreoni, L., Bartos, A., Berthele, A., Blankenstein, M. A., ... Teunissen, C. E. (2010). Neurofilament ELISA validation. Journal of Immunological Methods, 352(1-2), 23-31. https://doi.org/10.1016/j.jim.2009.09.014

Neurofilament ELISA validation. / Petzold, Axel; Altintas, Ayse; Andreoni, Laura; Bartos, Ales; Berthele, Achim; Blankenstein, Marinus A.; Buee, Luc; Castellazzi, Massimiliano; Cepok, Sabine; Comabella, Manuel; Constantinescu, Cris S.; Deisenhammer, Florian; Deniz, Gunnur; Erten, Gaye; Espiño, Mercedes; Fainardi, Enrico; Franciotta, Diego; Freedman, Mark S.; Giedraitis, Vilmantas; Gilhus, Nils Erik; Giovannoni, Gavin; Glabinski, Andrzej; Grieb, Pawel; Hartung, Hans Peter; Hemmer, Bernhard; Herukka, Sanna Kaisa; Hintzen, Rogier; Ingelsson, Martin; Jackson, Samuel; Jacobsen, Steve; Jafari, Naghmeh; Jalosinski, Marcin; Jarius, Sven; Kapaki, Elisabeth; Kieseier, Bernd C.; Koel-Simmelink, Marleen J A; Kornhuber, Johannes; Kuhle, Jens; Kurzepa, Jacek; Lalive, Patrice H.; Lannfelt, Lars; Lehmensiek, Vera; Lewczuk, Piotr; Livrea, Paolo; Marnetto, Fabiana; Martino, Davide; Menge, Til; Norgren, Niklas; Papuć, Eva; Paraskevas, George P.; Pirttilä, Tuula; Rajda, Cecília; Rejdak, Konrad; Ricny, Jan; Ripova, Daniela; Rosengren, Lars; Ruggieri, Maddalena; Schraen, Susanna; Shaw, Gerry; Sindic, Christian; Siva, Aksel; Stigbrand, Torgny; Stonebridge, Iva; Topcular, Baris; Trojano, Maria; Tumani, Hayrettin; Twaalfhoven, Harry A M; Vécsei, L.; Van Pesch, Vincent; Vanderstichele, Hugo; Vedeler, Christian; Verbeek, Marcel M.; Villar, Luisa Maria; Weissert, Robert; Wildemann, Brigitte; Yang, Cui; Yao, Karen; Teunissen, Charlotte E.

In: Journal of Immunological Methods, Vol. 352, No. 1-2, 31.01.2010, p. 23-31.

Research output: Contribution to journalArticle

Petzold, A, Altintas, A, Andreoni, L, Bartos, A, Berthele, A, Blankenstein, MA, Buee, L, Castellazzi, M, Cepok, S, Comabella, M, Constantinescu, CS, Deisenhammer, F, Deniz, G, Erten, G, Espiño, M, Fainardi, E, Franciotta, D, Freedman, MS, Giedraitis, V, Gilhus, NE, Giovannoni, G, Glabinski, A, Grieb, P, Hartung, HP, Hemmer, B, Herukka, SK, Hintzen, R, Ingelsson, M, Jackson, S, Jacobsen, S, Jafari, N, Jalosinski, M, Jarius, S, Kapaki, E, Kieseier, BC, Koel-Simmelink, MJA, Kornhuber, J, Kuhle, J, Kurzepa, J, Lalive, PH, Lannfelt, L, Lehmensiek, V, Lewczuk, P, Livrea, P, Marnetto, F, Martino, D, Menge, T, Norgren, N, Papuć, E, Paraskevas, GP, Pirttilä, T, Rajda, C, Rejdak, K, Ricny, J, Ripova, D, Rosengren, L, Ruggieri, M, Schraen, S, Shaw, G, Sindic, C, Siva, A, Stigbrand, T, Stonebridge, I, Topcular, B, Trojano, M, Tumani, H, Twaalfhoven, HAM, Vécsei, L, Van Pesch, V, Vanderstichele, H, Vedeler, C, Verbeek, MM, Villar, LM, Weissert, R, Wildemann, B, Yang, C, Yao, K & Teunissen, CE 2010, 'Neurofilament ELISA validation', Journal of Immunological Methods, vol. 352, no. 1-2, pp. 23-31. https://doi.org/10.1016/j.jim.2009.09.014
Petzold A, Altintas A, Andreoni L, Bartos A, Berthele A, Blankenstein MA et al. Neurofilament ELISA validation. Journal of Immunological Methods. 2010 Jan 31;352(1-2):23-31. https://doi.org/10.1016/j.jim.2009.09.014
Petzold, Axel ; Altintas, Ayse ; Andreoni, Laura ; Bartos, Ales ; Berthele, Achim ; Blankenstein, Marinus A. ; Buee, Luc ; Castellazzi, Massimiliano ; Cepok, Sabine ; Comabella, Manuel ; Constantinescu, Cris S. ; Deisenhammer, Florian ; Deniz, Gunnur ; Erten, Gaye ; Espiño, Mercedes ; Fainardi, Enrico ; Franciotta, Diego ; Freedman, Mark S. ; Giedraitis, Vilmantas ; Gilhus, Nils Erik ; Giovannoni, Gavin ; Glabinski, Andrzej ; Grieb, Pawel ; Hartung, Hans Peter ; Hemmer, Bernhard ; Herukka, Sanna Kaisa ; Hintzen, Rogier ; Ingelsson, Martin ; Jackson, Samuel ; Jacobsen, Steve ; Jafari, Naghmeh ; Jalosinski, Marcin ; Jarius, Sven ; Kapaki, Elisabeth ; Kieseier, Bernd C. ; Koel-Simmelink, Marleen J A ; Kornhuber, Johannes ; Kuhle, Jens ; Kurzepa, Jacek ; Lalive, Patrice H. ; Lannfelt, Lars ; Lehmensiek, Vera ; Lewczuk, Piotr ; Livrea, Paolo ; Marnetto, Fabiana ; Martino, Davide ; Menge, Til ; Norgren, Niklas ; Papuć, Eva ; Paraskevas, George P. ; Pirttilä, Tuula ; Rajda, Cecília ; Rejdak, Konrad ; Ricny, Jan ; Ripova, Daniela ; Rosengren, Lars ; Ruggieri, Maddalena ; Schraen, Susanna ; Shaw, Gerry ; Sindic, Christian ; Siva, Aksel ; Stigbrand, Torgny ; Stonebridge, Iva ; Topcular, Baris ; Trojano, Maria ; Tumani, Hayrettin ; Twaalfhoven, Harry A M ; Vécsei, L. ; Van Pesch, Vincent ; Vanderstichele, Hugo ; Vedeler, Christian ; Verbeek, Marcel M. ; Villar, Luisa Maria ; Weissert, Robert ; Wildemann, Brigitte ; Yang, Cui ; Yao, Karen ; Teunissen, Charlotte E. / Neurofilament ELISA validation. In: Journal of Immunological Methods. 2010 ; Vol. 352, No. 1-2. pp. 23-31.
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abstract = "Background: Neurofilament proteins (Nf) are highly specific biomarkers for neuronal death and axonal degeneration. As these markers become more widely used, an inter-laboratory validation study is required to identify assay criteria for high quality performance. Methods: The UmanDiagnostics NF-light {\circledR}enzyme-linked immunoabsorbent assays (ELISA) for the neurofilament light chain (NfL, 68 kDa) was used to test the intra-assay and inter-laboratory coefficient of variation (CV) between 35 laboratories worldwide on 15 cerebrospinal fluid (CSF) samples. Critical factors, such as sample transport and storage, analytical delays, reaction temperature and time, the laboratories' accuracy and preparation of standards were documented and used for the statistical analyses. Results: The intra-laboratory CV averaged 3.3{\%} and the inter-laboratory CV 59{\%}. The results from the test laboratories correlated with those from the reference laboratory (R = 0.60, p <0.0001). Correcting for critical factors improved the strength of the correlation. Differences in the accuracy of standard preparation were identified as the most critical factor. Correcting for the error introduced by variation in the protein standards improved the correlation to R = 0.98, p <0.0001 with an averaged inter-laboratory CV of 14{\%}. The corrected overall inter-rater agreement was subtantial (0.6) according to Fleiss' multi-rater kappa and Gwet's AC1 statistics. Conclusion: This multi-center validation study identified the lack of preparation of accurate and consistent protein standards as the main reason for a poor inter-laboratory CV. This issue is also relevant to other protein biomarkers based on this type of assay and will need to be solved in order to achieve an acceptable level of analytical accuracy. The raw data of this study is available online.",
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TY - JOUR

T1 - Neurofilament ELISA validation

AU - Petzold, Axel

AU - Altintas, Ayse

AU - Andreoni, Laura

AU - Bartos, Ales

AU - Berthele, Achim

AU - Blankenstein, Marinus A.

AU - Buee, Luc

AU - Castellazzi, Massimiliano

AU - Cepok, Sabine

AU - Comabella, Manuel

AU - Constantinescu, Cris S.

AU - Deisenhammer, Florian

AU - Deniz, Gunnur

AU - Erten, Gaye

AU - Espiño, Mercedes

AU - Fainardi, Enrico

AU - Franciotta, Diego

AU - Freedman, Mark S.

AU - Giedraitis, Vilmantas

AU - Gilhus, Nils Erik

AU - Giovannoni, Gavin

AU - Glabinski, Andrzej

AU - Grieb, Pawel

AU - Hartung, Hans Peter

AU - Hemmer, Bernhard

AU - Herukka, Sanna Kaisa

AU - Hintzen, Rogier

AU - Ingelsson, Martin

AU - Jackson, Samuel

AU - Jacobsen, Steve

AU - Jafari, Naghmeh

AU - Jalosinski, Marcin

AU - Jarius, Sven

AU - Kapaki, Elisabeth

AU - Kieseier, Bernd C.

AU - Koel-Simmelink, Marleen J A

AU - Kornhuber, Johannes

AU - Kuhle, Jens

AU - Kurzepa, Jacek

AU - Lalive, Patrice H.

AU - Lannfelt, Lars

AU - Lehmensiek, Vera

AU - Lewczuk, Piotr

AU - Livrea, Paolo

AU - Marnetto, Fabiana

AU - Martino, Davide

AU - Menge, Til

AU - Norgren, Niklas

AU - Papuć, Eva

AU - Paraskevas, George P.

AU - Pirttilä, Tuula

AU - Rajda, Cecília

AU - Rejdak, Konrad

AU - Ricny, Jan

AU - Ripova, Daniela

AU - Rosengren, Lars

AU - Ruggieri, Maddalena

AU - Schraen, Susanna

AU - Shaw, Gerry

AU - Sindic, Christian

AU - Siva, Aksel

AU - Stigbrand, Torgny

AU - Stonebridge, Iva

AU - Topcular, Baris

AU - Trojano, Maria

AU - Tumani, Hayrettin

AU - Twaalfhoven, Harry A M

AU - Vécsei, L.

AU - Van Pesch, Vincent

AU - Vanderstichele, Hugo

AU - Vedeler, Christian

AU - Verbeek, Marcel M.

AU - Villar, Luisa Maria

AU - Weissert, Robert

AU - Wildemann, Brigitte

AU - Yang, Cui

AU - Yao, Karen

AU - Teunissen, Charlotte E.

PY - 2010/1/31

Y1 - 2010/1/31

N2 - Background: Neurofilament proteins (Nf) are highly specific biomarkers for neuronal death and axonal degeneration. As these markers become more widely used, an inter-laboratory validation study is required to identify assay criteria for high quality performance. Methods: The UmanDiagnostics NF-light ®enzyme-linked immunoabsorbent assays (ELISA) for the neurofilament light chain (NfL, 68 kDa) was used to test the intra-assay and inter-laboratory coefficient of variation (CV) between 35 laboratories worldwide on 15 cerebrospinal fluid (CSF) samples. Critical factors, such as sample transport and storage, analytical delays, reaction temperature and time, the laboratories' accuracy and preparation of standards were documented and used for the statistical analyses. Results: The intra-laboratory CV averaged 3.3% and the inter-laboratory CV 59%. The results from the test laboratories correlated with those from the reference laboratory (R = 0.60, p <0.0001). Correcting for critical factors improved the strength of the correlation. Differences in the accuracy of standard preparation were identified as the most critical factor. Correcting for the error introduced by variation in the protein standards improved the correlation to R = 0.98, p <0.0001 with an averaged inter-laboratory CV of 14%. The corrected overall inter-rater agreement was subtantial (0.6) according to Fleiss' multi-rater kappa and Gwet's AC1 statistics. Conclusion: This multi-center validation study identified the lack of preparation of accurate and consistent protein standards as the main reason for a poor inter-laboratory CV. This issue is also relevant to other protein biomarkers based on this type of assay and will need to be solved in order to achieve an acceptable level of analytical accuracy. The raw data of this study is available online.

AB - Background: Neurofilament proteins (Nf) are highly specific biomarkers for neuronal death and axonal degeneration. As these markers become more widely used, an inter-laboratory validation study is required to identify assay criteria for high quality performance. Methods: The UmanDiagnostics NF-light ®enzyme-linked immunoabsorbent assays (ELISA) for the neurofilament light chain (NfL, 68 kDa) was used to test the intra-assay and inter-laboratory coefficient of variation (CV) between 35 laboratories worldwide on 15 cerebrospinal fluid (CSF) samples. Critical factors, such as sample transport and storage, analytical delays, reaction temperature and time, the laboratories' accuracy and preparation of standards were documented and used for the statistical analyses. Results: The intra-laboratory CV averaged 3.3% and the inter-laboratory CV 59%. The results from the test laboratories correlated with those from the reference laboratory (R = 0.60, p <0.0001). Correcting for critical factors improved the strength of the correlation. Differences in the accuracy of standard preparation were identified as the most critical factor. Correcting for the error introduced by variation in the protein standards improved the correlation to R = 0.98, p <0.0001 with an averaged inter-laboratory CV of 14%. The corrected overall inter-rater agreement was subtantial (0.6) according to Fleiss' multi-rater kappa and Gwet's AC1 statistics. Conclusion: This multi-center validation study identified the lack of preparation of accurate and consistent protein standards as the main reason for a poor inter-laboratory CV. This issue is also relevant to other protein biomarkers based on this type of assay and will need to be solved in order to achieve an acceptable level of analytical accuracy. The raw data of this study is available online.

KW - Biomarker

KW - NEFL

KW - Neurofilament

KW - NF-L

KW - NfL

KW - Validation

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