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An Improved Assay for Quantitation of Cerebrospinal Fluid Cystatin C Using Liquid Chromatography Tandem Mass Spectrometry

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Cerebrospinal Fluid (CSF) Proteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2044))

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Abstract

Cystatin C (CST3) is expressed ubiquitously and implicated in several neurological diseases. It can be posttranscriptionally modified. CST3 is usually quantified in a biological sample using antibody-based methods. Posttranscriptional modification can hamper antibody-based detection systems by altering antibody-binding epitope(s). To circumvent this problem, enzymatic digestion and liquid chromatography tandem mass spectrometry (LC-MS/MS) technique can be employed to identify and measure peptides of a target protein in a complex biological mixture. This chapter describes an LC-MS/MS-based method for accurate measurement of CST3 in cerebrospinal fluid (CSF). Here, CSF was directly subjected to trypsin digestion and digested peptides were extracted using a solid-phase extraction column. Extracted peptide samples were directly used for LC-MS/MS-based identification and quantification of CST3 peptides. Comparing the concentration in a set of samples measured by LC-MS/MS with that of immunoassay shows that it was significantly higher when measured by LC-MS/MS method, suggesting it a better quantification method. This approach is particularly well suited when posttranscriptional modification of CST3 is suspected and sample volume of CSF is small.

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Authors and Affiliations

  1. Department of Laboratory Medicine, Faculty of Medicine, Shimane University, Izumo, Japan

    Abdullah Md Sheikh & Atsushi Nagai

Authors
  1. Abdullah Md Sheikh
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  2. Atsushi Nagai
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Corresponding author

Correspondence to Atsushi Nagai .

Editor information

Editors and Affiliations

  1. Proteomics Unit, Clinical Neuroproteomics Laboratory, Navarrabiomed, Complejo Hospitalario de Navarra (CHN), Universidad Pública de Navarra (UPNA), IdiSNA, Proteored-ISCIII, Pamplona, Spain

    Enrique Santamaría

  2. Proteomics Unit, Clinical Neuroproteomics Laboratory, Navarrabiomed, Complejo Hospitalario de Navarra (CHN), Universidad Pública de Navarra (UPNA), IdiSNA, Proteored-ISCIII, Pamplona, Spain

    Joaquín Fernández-Irigoyen

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Sheikh, A.M., Nagai, A. (2019). An Improved Assay for Quantitation of Cerebrospinal Fluid Cystatin C Using Liquid Chromatography Tandem Mass Spectrometry. In: Santamaría, E., Fernández-Irigoyen, J. (eds) Cerebrospinal Fluid (CSF) Proteomics. Methods in Molecular Biology, vol 2044. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9706-0_18

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  • DOI: https://doi.org/10.1007/978-1-4939-9706-0_18

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9705-3

  • Online ISBN: 978-1-4939-9706-0

  • eBook Packages: Springer Protocols

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