Development and Validation of an Ultra-Performance Liquid Chromatography with Tandem Mass Spectrometry Method for Determination of Soluble Repulsive Guidance Molecule A in Human Serum and Cerebrospinal Fluid

Patrick Schreier,Lili Huang,Emma Fung,Jennifer Mollon,Bernhard Sielaff,Marc R. Lake,Michael Schulz,Khader Awwad

Bioanalysis（2024）SCI 4区SCI 3区

AbbVie Deutschland GmbH & Co KG | AbbVie Biores Ctr | AbbVie Inc

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Abstract

Aim: Repulsive guidance molecule A (RGMa) is upregulated in neurodegenerative diseases. To assess RGMa levels in human serum and cerebrospinal fluid (CSF), a quantification method was developed and validated according to ICH M10 guideline. Methods & results: Sample preparation consisted of immunoprecipitation (IP, only for serum), digestion and purification followed by MS. Conclusion: An UPLC-MS/MS method was established and used to assess normal range of soluble RGMa levels in serum and CSF of healthy controls, and patients with mild cognitive impairment or Alzheimer's disease. The normal range was between 13.0-44.8 ng/ml (CSF) and 9.9-20.9 ng/ml (serum) in healthy controls. In the CSF of patients with mild cognitive impairment and Alzheimer's disease, total soluble RGMa was twofold lower while unchanged in serum.

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Key words

biomarker,immunoprecipitation,method development/validation according to ICH M10 guideline,neurodegeneration,soluble RGMa,UPLC-MS/MS

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