Targeted human cerebrospinal fluid proteomics for the validation of multiple Alzheimerʹs disease biomarker candidates

There is significant interest in the development of methods to validate novel biomarkers for Alzheimerʹs disease (AD) diagnosis. Previously, a proteomic panel of cerebrospinal fluid (CSF) biomarker candidates that differentiated AD and non-AD CSF with accuracy higher than 90% was found; information about these CSF proteins can be used to develop multiple reaction monitoring (MRM) based analytical assays, which offer the possibility of quantifying protein expression level changes in samples, as well as, validation among multiple laboratories. Here we report an MRM assay that demonstrates good linearity (average R2 = 0.969) and reproducibility (average coefficient of variance of 6.93%) for the proposed AD CSF biomarkers. MRM quantification results of Aβ1-40, Aβ1-42, retinol-binding protein and cystatin C correlated well with those from ELISA (average R2 = 0.974). Analysis shows that 12 out of 16 selected targets exhibit the same trend in protein expression as that in literature.