Development of a Highly Sensitive Neurofilament Light Chain Assay on an Automated Immunoassay Platform
BackgroundNeurofilament light chain (NfL) is an axonal cytoskeletal protein that is released into the extracellular space following neuronal or axonal injury associated with neurological conditions such as multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS), and other diseases. NfL is detec...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-07-01
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| Series: | Frontiers in Neurology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fneur.2022.935382/full |
| _version_ | 1817969196107038720 |
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| author | Stephen Lee Tatiana Plavina Carol M. Singh Kuangnan Xiong Xiaolei Qiu Richard A. Rudick Peter A. Calabresi Lauren Stevenson Danielle Graham Denitza Raitcheva Christopher Green Madeleine Matias Arejas J. Uzgiris |
| author_facet | Stephen Lee Tatiana Plavina Carol M. Singh Kuangnan Xiong Xiaolei Qiu Richard A. Rudick Peter A. Calabresi Lauren Stevenson Danielle Graham Denitza Raitcheva Christopher Green Madeleine Matias Arejas J. Uzgiris |
| author_sort | Stephen Lee |
| collection | DOAJ |
| description | BackgroundNeurofilament light chain (NfL) is an axonal cytoskeletal protein that is released into the extracellular space following neuronal or axonal injury associated with neurological conditions such as multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS), and other diseases. NfL is detectable in the cerebrospinal fluid (CSF) and blood. Numerous studies on MS have demonstrated that NfL is correlated with disease activity, predicts disease progression, and is reduced by treatment with MS disease-modifying drugs, making NfL an attractive candidate to supplement existing clinical and imaging measures in MS. However, for NfL to achieve its potential as a clinically useful biomarker for clinical decision-making or drug development, a standardized, practical, and widely accessible assay is needed. Our objective was to develop a novel NfL assay on an automated, globally available immunoassay platform and validate its performance.MethodsA prototype NfL assay was first developed and evaluated on the ADVIA Centaur® XP immunoassay system from Siemens Healthineers. The lower limit of quantitation (LLoQ), within-lab precision, assay range, cross-reactivity with neurofilament medium and heavy chains, and effect of interfering substances were determined. NfL assay values in serum and CSF were compared with radiological and clinical disease activity measures in patients with MS and ALS, respectively. This assay was further optimized to utilize serum, plasma, and CSF sample types on the Atellica® IM system and transferred to Siemens' CLIA laboratory where it was analytically validated as a laboratory-developed test (LDT).ResultsIn this study, an LLoQ of 1.85 pg/mL, within-lab precision <6%, and an assay range of up to 646 pg/mL were demonstrated with the serum prototype assay. Cross-reactivity of <0.7% with the neurofilament medium and heavy chains was observed. Serum and CSF NfL assay values were associated with radiological and clinical disease activity measures in patients with MS and ALS, respectively. The optimized version of the NfL assay demonstrated specimen equivalence with additional plasma tube types and was analytically validated as an LDT.ConclusionThe analytical performance of the NfL assay fulfilled all acceptance criteria; therefore, we suggest that the assay is acceptable for use in both research and clinical practice settings to determine elevated NfL levels in patients. |
| first_indexed | 2024-04-13T20:18:16Z |
| format | Article |
| id | doaj.art-7eb04dd5b6fb446fb7e55dba0540b672 |
| institution | Directory Open Access Journal |
| issn | 1664-2295 |
| language | English |
| last_indexed | 2024-04-13T20:18:16Z |
| publishDate | 2022-07-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Neurology |
| spelling | doaj.art-7eb04dd5b6fb446fb7e55dba0540b6722022-12-22T02:31:37ZengFrontiers Media S.A.Frontiers in Neurology1664-22952022-07-011310.3389/fneur.2022.935382935382Development of a Highly Sensitive Neurofilament Light Chain Assay on an Automated Immunoassay PlatformStephen Lee0Tatiana Plavina1Carol M. Singh2Kuangnan Xiong3Xiaolei Qiu4Richard A. Rudick5Peter A. Calabresi6Lauren Stevenson7Danielle Graham8Denitza Raitcheva9Christopher Green10Madeleine Matias11Arejas J. Uzgiris12Siemens Healthcare Laboratory, LLC, Berkeley, CA, United StatesBiogen, Cambridge, MA, United StatesBiogen, Cambridge, MA, United StatesBiogen, Cambridge, MA, United StatesSiemens Healthcare Laboratory, LLC, Berkeley, CA, United StatesBiogen, Cambridge, MA, United StatesDepartment of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD, United StatesBiogen, Cambridge, MA, United StatesBiogen, Cambridge, MA, United StatesBiogen, Cambridge, MA, United StatesSiemens Healthcare Laboratory, LLC, Berkeley, CA, United StatesSiemens Healthcare Laboratory, LLC, Berkeley, CA, United StatesSiemens Healthcare Laboratory, LLC, Berkeley, CA, United StatesBackgroundNeurofilament light chain (NfL) is an axonal cytoskeletal protein that is released into the extracellular space following neuronal or axonal injury associated with neurological conditions such as multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS), and other diseases. NfL is detectable in the cerebrospinal fluid (CSF) and blood. Numerous studies on MS have demonstrated that NfL is correlated with disease activity, predicts disease progression, and is reduced by treatment with MS disease-modifying drugs, making NfL an attractive candidate to supplement existing clinical and imaging measures in MS. However, for NfL to achieve its potential as a clinically useful biomarker for clinical decision-making or drug development, a standardized, practical, and widely accessible assay is needed. Our objective was to develop a novel NfL assay on an automated, globally available immunoassay platform and validate its performance.MethodsA prototype NfL assay was first developed and evaluated on the ADVIA Centaur® XP immunoassay system from Siemens Healthineers. The lower limit of quantitation (LLoQ), within-lab precision, assay range, cross-reactivity with neurofilament medium and heavy chains, and effect of interfering substances were determined. NfL assay values in serum and CSF were compared with radiological and clinical disease activity measures in patients with MS and ALS, respectively. This assay was further optimized to utilize serum, plasma, and CSF sample types on the Atellica® IM system and transferred to Siemens' CLIA laboratory where it was analytically validated as a laboratory-developed test (LDT).ResultsIn this study, an LLoQ of 1.85 pg/mL, within-lab precision <6%, and an assay range of up to 646 pg/mL were demonstrated with the serum prototype assay. Cross-reactivity of <0.7% with the neurofilament medium and heavy chains was observed. Serum and CSF NfL assay values were associated with radiological and clinical disease activity measures in patients with MS and ALS, respectively. The optimized version of the NfL assay demonstrated specimen equivalence with additional plasma tube types and was analytically validated as an LDT.ConclusionThe analytical performance of the NfL assay fulfilled all acceptance criteria; therefore, we suggest that the assay is acceptable for use in both research and clinical practice settings to determine elevated NfL levels in patients.https://www.frontiersin.org/articles/10.3389/fneur.2022.935382/fullassayneurofilamentbiomarkerneurodegenerative article type: original research manuscript sectionmultiple sclerosisamyotrophic lateral sclerosis |
| spellingShingle | Stephen Lee Tatiana Plavina Carol M. Singh Kuangnan Xiong Xiaolei Qiu Richard A. Rudick Peter A. Calabresi Lauren Stevenson Danielle Graham Denitza Raitcheva Christopher Green Madeleine Matias Arejas J. Uzgiris Development of a Highly Sensitive Neurofilament Light Chain Assay on an Automated Immunoassay Platform Frontiers in Neurology assay neurofilament biomarker neurodegenerative article type: original research manuscript section multiple sclerosis amyotrophic lateral sclerosis |
| title | Development of a Highly Sensitive Neurofilament Light Chain Assay on an Automated Immunoassay Platform |
| title_full | Development of a Highly Sensitive Neurofilament Light Chain Assay on an Automated Immunoassay Platform |
| title_fullStr | Development of a Highly Sensitive Neurofilament Light Chain Assay on an Automated Immunoassay Platform |
| title_full_unstemmed | Development of a Highly Sensitive Neurofilament Light Chain Assay on an Automated Immunoassay Platform |
| title_short | Development of a Highly Sensitive Neurofilament Light Chain Assay on an Automated Immunoassay Platform |
| title_sort | development of a highly sensitive neurofilament light chain assay on an automated immunoassay platform |
| topic | assay neurofilament biomarker neurodegenerative article type: original research manuscript section multiple sclerosis amyotrophic lateral sclerosis |
| url | https://www.frontiersin.org/articles/10.3389/fneur.2022.935382/full |
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