Assessment of immunoprecipitation with subsequent immunoassays for the blood-based diagnosis of Alzheimer’s disease

Abstract

The Aβ42/40 ratio and the concentration of phosphorylated Tau181 in blood plasma represent attractive biomarkers for Alzheimer's disease. As a means for reducing potential matrix effects, which may interfere with plasma immunoassays, we have previously developed a pre-analytical sample workup by semi-automated immunoprecipitation. Here we test the compatibility of pre-analytical immunoprecipitations with automated Aβ1-40, Aβ1-42 and phosphorylated Tau181 immunoassays on the Lumipulse platform and compare the diagnostic performance of the respective immunoprecipitation immunoassay approaches with direct plasma measurements. 71 participants were dichotomized according to their Aβ42/40 ratios in cerebrospinal fluid into the diagnostic groups amyloid-positive (n = 32) and amyloid-negative (n = 39). The plasma Aβ1-42/1-40 ratio and phosphorylated Tau181 levels were determined on the Lumipulse G600II platform (Fujirebio) by direct measurements in EDTA–plasma or after Aβ- or Tau-immunoprecipitation, respectively. Pre-analytical immunoprecipitation of Aβ turned out to be compatible with the Lumipulse Aβ assays and resulted in a numerical, yet statistically not significant increase in the area under the ROC curve for plasma Aβ1-42/1-40. Additionally, we observed a significant increase in the standardised effect size (Cohen’s D). Pre-analytical immunoprecipitation of Tau resulted in increased differences between the diagnostic groups in terms of median and mean phosphorylated Tau 181 levels. Furthermore, we observed a greater Cohen’s d (p < 0.001) and a larger area under the ROC curve (p = 0.038) after Tau-IP. Our preliminary findings in a small, preselected sample indicate that pre-analytical immunoprecipitation may have the potential to improve the diagnostic performance of plasma biomarker immunoassays for Aβ1-42/1-40 and phosphorylated Tau181 to predict brain amyloid deposition.