An automated clinical mass spectrometric method for identification and quantification of variant and wild-type amyloid-ß 1-40 and 1-42 peptides in CSF.

ABSTRACT

INTRODUCTION: We developed an automated liquid chromatography-tandem mass spectrometry high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method for multiplex quantification of wild-type (wt) amyloid ß (Aß) peptides 1-40 (Aß40) and 1-42 (Aß42) and detection of variant Aß peptides in cerebrospinal fluid. METHODS: The multiplex Aß HPLC-MS/MS assay was validated in a clinically accredited laboratory following regulatory guidelines, with Aß42 calibration assigned to the ERM/IFCC certified reference material; sequence variants were additionally multiplexed into the method. RESULTS: Sample preparation was fully automated on a liquid handler. The assay quantified wt-Aß42 and wt-Aß40 and detected sequence variants, when present, within the Aß42 sequence. DISCUSSION: Extension of the HPLC-MS/MS approach for quantification of wt-Aß42 and wt-Aß40 to include known sequence variants increases analytical accuracy of the mass spectrometric approach and enables identification of cases of autosomal dominant Alzheimer's disease. Development of an automated workflow and selection of appropriate instrumentation enabled deployment of this method in routine clinical testing.