Antibody-free detection of amyloid beta peptides biomarkers in cerebrospinal fluid using capillary isotachophoresis coupled with mass spectrometry

Abstract

This study reports a capillary isotachophoresis (ITP) – electrospray ionization mass spectrometry (ESI-MS) method for the determination of several amyloid β (Aβ) peptides, which are biomarkers of Alzheimer’s disease (AD) in cerebrospinal fluids (CSF). For the first time, these peptides have been detected directly from CSF by MS without recourse to an immunocapture-based sample pre-treatment. The antibody-free approach is based on the marriage between capillary ITP, a powerful on-line electrokinetic preconcentration technique, and MS for simultaneous detection of different Aβ peptides. To ensure a good performance, the ITP process of fluorescently labelled Aβ peptides was for the first time implemented and verified with laser induced fluorescent detection, prior to methodology transfer to MS detection. Better detection sensitivity was achieved with labelled Aβ peptides for both detection modes. Using hydroxyl ions as the terminating and acetate as the leading ions, our method allows efficient ITP preconcentration under alkaline conditions of the slowly migrating Aβ peptides to reach quantifiable concentration down to 50 pM. The developed ITP-MS approach allows reliable quantification of different fluorescently derivatized Aβ peptides, i.e. Aβ 1–42, Aβ 1–40 and Aβ 1–38 down to sub nM ranges in CSF samples from AD and non-demented (healthy control) patients. Good agreement (<20% deviation) for the determination of Aβ 1–42/Aβ 1–40 ratio in CSF was achieved between results obtained with this new ITP-MS and our recently developed method based on large volume sample stacking coupled to CE. Discrimination of one AD patient from two healthy controls was successfully made with the Aβ 1–42/Aβ 1–40 ratio obtained by the developed ITP-MS method for the tested cerebrospinal fluid samples.