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An integrated microfluidic chip for immunocapture, preconcentration and separation of β-amyloid peptides

Authors: Mohamadi M. Reza | Svobodová Zuzana | Bílková Zuzana | Otto Markus | Taverna Myriam | Descroix Stephanie | Viovy Jean-Louis

Year: 2015

Type of publication: článek v odborném periodiku

Name of source: Biomicrofluidics

Page from-to: 054117

Titles:

Language	Name	Abstract	Keywords
cze	Integrované mikrofluidní zařízení pro záchyt, prekoncentraci a separaci peptidů amyloidu beta	Ve článku je prezentováno integrované mikrofluidní zařízení pro detekci peptidů amyloidu beta. Tyto peptidy se řadí mezi hlavní biomarkery při diagnostice Alzheimerovy choroby (AD) v časné fázi onemocnění. Toto mikrofluidní zařízení se skládá ze tří hlavních komponent: (1) mikrokolonky pro záchyt peptidů amyloidu beta na magnetických částicích za pomoci specifických protilátek, (2) nanoporézní membrány vyrobené z fotopolymerizovatelného hydrogelu pro prekoncentraci izolovaných peptidů a (3) mikročipové elektroforézy s fluorescenční detekcí. K detekci stačí 25 ng syntetického peptidu v mozkomišním moku. Analýza byla vyzkoušena na zdravých kontrolách i na pacientech s frontotemporální demecí nebo AD. Zařízení umožňuje detekovat následující izoformy peptidů amyloidu beta (1–37, 1–39, 1–40, and 1–42).	Mikrofluidika; integrované zařízení; kapilární elektroforéza; hydrogelová membrána; amyloid beta; peptide; Alzheimerova choroba; frontotemporální choroba
eng	An integrated microfluidic chip for immunocapture, preconcentration and separation of β-amyloid peptides	We present an integrated microfluidic chip for detection of β-amyloid (Aβ) peptides. Aβ peptides are major biomarkers for the diagnosis of Alzheimer's disease (AD) in its early stages. This microfluidic device consists of three main parts: (1) An immunocapture microcolumn based on self-assembled magnetic beads coated with antibodies specific to Aβ peptides, (2) a nano-porous membrane made of photopolymerized hydrogel for preconcentration, and (3) a microchip electrophoresis (MCE) channel with fluorescent detection. Sub-milliliter sample volume is either mixed off-chip with antibody coated magnetic beads and injected into the device or is injected into an already self-assembled column of magnetic beads in the microchannel. The captured peptides on the beads are then electrokinetically eluted and re-concentrated onto the nano-membrane in a few nano-liters. By integrating the nano-membrane, total assay time was reduced and also off-chip re-concentration or buffer exchange steps were not needed. Finally, the concentrated peptides in the chip are separated by electrophoresis in a polymer-based matrix. The device was applied to the capture and MCE analysis of differently truncated peptides Aβ (1–37, 1–39, 1–40, and 1–42) and was able to detect as low as 25 ng of synthetic Aβ peptides spiked in undiluted cerebrospinal fluid (CSF). The device was also tested with CSF samples from healthy donors. CSF samples were fluorescently labelled and pre-mixed with the magnetic beads and injected into the device. The results indicated that Aβ1-40, an important biomarker for distinguishing patients with frontotemporal lobe dementia from controls and AD patients, was detectable. Although the sensitivity of this device is not yet enough to detect all Aβ subtypes in CSF, this is the first report on an integrated or semi-integrated device for capturing and analyzing of differently truncated Aβ peptides.	Microfluidic; integrated device; capillary electrophoresis; hydrogel membrane; amyloid beta; peptide; Alzheimer disease; Frontotemporal disease