Gelsolin bound beta-amyloid peptides((1-40/1-42)): Electrochemical evaluation of levels of soluble peptide associated with Alzheimer's disease

摘要

A method for the highly sensitive determination of soluable beta-amyloid peptides (A beta(1-40/1-42)) that employs a detection bioconjugate of HRP-Au-gelsolin as the electrochemical nanoprobe is presented. Contrary to previous detection notions that utilized antibodies, which could specifically recognize the N- or C-terminus of peptides, we demonstrate herein that the reported specific binding between gelsolin and A beta might provide an alternative way to evaluate the peptides sensitively and selectively. The HRP-Au-gelsolin nanohybrid was designed by one-pot functionalization of Au nanaoparticles (NPs) with horseradish peroxidase (HRP) and gelsolin. Through a sandwich-type sensor array, soluble A beta(1-40)/(1-42) were captured onto the array due to the interactions between targeted peptides and surface-confined gelsolin and electrochemical signals were amplified by abundant attachments of HRP labeled on AuNPs, which could specifically catalyse its substrate, 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2 to give rise to measurable signals. The proposed gelsolin-bound A beta methodology displayed satisfactory sensitivity and wide linear range towards A beta(1-40/1-42) with a detection limit down to 28 pM, which are verified to be sensitive-enough for the assessment of A beta levels both in normal and Alzheimer's disease (AD) rat brains. Experimental results indicated that compared with normal group, soluble beta-amyloid peptide levels in cerebrospinal fluid (CSF) and targeted brain tissues of AD rats all declined with differentiable degrees. In short, the newly unfolding strategy presents valuable information related to pathological events in brain and will exhibit a braw perspective for the early diagnosis of AD process. (C) 2014 Elsevier B.V. All rights reserved.