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Mass spectrometric analysis of nanoscale sample volumes extracted from open microchannels after sample preconcentration applied on amyloid beta peptides
KTH, Skolan för kemivetenskap (CHE), Kemi, Tillämpad fysikalisk kemi.
KTH, Skolan för kemivetenskap (CHE), Kemi, Tillämpad fysikalisk kemi.ORCID-id: 0000-0003-3548-217X
KTH, Skolan för kemivetenskap (CHE), Kemi, Tillämpad fysikalisk kemi.ORCID-id: 0000-0002-3444-9987
2014 (Engelska)Ingår i: Analytical and Bioanalytical Chemistry, ISSN 1618-2642, E-ISSN 1618-2650, Vol. 406, nr 14, s. 3521-3524Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

A new instrumental concept for extraction of nanovolumes from open microchannels (dimensions 150 mu m x 50 mu m, length 10 mm) manufactured on silicon microchips has been used in combination with a previously developed method for preconcentrating proteins and peptides in the open channels through electromigration. The extracted nanovolumes were further analyzed using nanoelectrospray ionization (nESI) or matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) directly or with subsequent enzymatic protein digestion in a nanodroplet prior to the MS analysis. Preconcentration of the samples resulted in a 15-fold sensitivity increase in nESI for a neurotensin solution, and using MALDI-MS, amyloid beta (A beta) peptides could be detected in concentrations down to 1 nM. The method was also successfully applied for detection of cell culture A beta.

Ort, förlag, år, upplaga, sidor
2014. Vol. 406, nr 14, s. 3521-3524
Nyckelord [en]
Mass spectrometry, Microchannel, Preconcentration, Amyloid beta, MALDI, Nano-ESI
Nationell ämneskategori
Biokemi och molekylärbiologi
Identifikatorer
URN: urn:nbn:se:kth:diva-147047DOI: 10.1007/s00216-014-7781-0ISI: 000336261800030Scopus ID: 2-s2.0-84901603229OAI: oai:DiVA.org:kth-147047DiVA, id: diva2:728330
Forskningsfinansiär
Vetenskapsrådet, 621-2009-4095
Anmärkning

QC 20140624

Tillgänglig från: 2014-06-24 Skapad: 2014-06-23 Senast uppdaterad: 2017-12-05Bibliografiskt granskad
Ingår i avhandling
1. Electrophoretic focusing in microchannels combined with mass spectrometry: Applications on amyloid beta peptides
Öppna denna publikation i ny flik eller fönster >>Electrophoretic focusing in microchannels combined with mass spectrometry: Applications on amyloid beta peptides
2016 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Abstract [en]

Analysis of low-abundance components in small samples remains a challenge within bioanalytical chemistry, and new techniques for sample pretreatments followed by sensitive and informative detection are required. In this thesis, procedures for preconcentration and separation of proteins and peptides in open microchannels fabricated on silicon microchips are presented. Analyte electromigration was induced by applying a voltage along the channel length, and detection was performed either by matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) within the open channel, or by sampling a nL fraction containing the preconcentrated analytes from the channel for subsequent nano-electrospray ionization- (nESI-) or MALDI-MS. Utilizing solvent evaporation from the open system during sample supply, sample volumes exceeding the 25-75 nL channel volume could be analyzed. For preconcentration/separation of components in the discrete channel volume a lid of inert fluorocarbon liquid was used for evaporation control.

In Papers I and II, aqueous, carrier-free solutions of proteins and peptides were analyzed, and the method was successfully applied for fast and simple preconcentration of amyloid beta (Aβ) peptides, related to Alzheimer’s disease.

The impact of possible impurities in the analysis of carrier-free solutions was investigated in Paper III with the 1D simulation software GENTRANS, and a method for open-channel isoelectric focusing in a tailor-made pH gradient was developed. The latter approach was used in Paper IV for preconcentration and purification of Aβ peptides after immunoprecipitation from cerebrospinal fluid and blood plasma, followed by MALDI-MS from a micropillar chip.

Paper V includes simulations of an isotachophoretic strategy for selective enrichment of Aβ peptides. GENTRANS simulations were used to select the electrolyte composition, and 2D simulations in a geometry suitable for on-chip implementation were performed using COMSOL Multiphysics.

Ort, förlag, år, upplaga, sidor
Stockholm: KTH Royal Institute of Technology, 2016. s. 55
Serie
TRITA-CHE-Report, ISSN 1654-1081 ; 2016:36
Nyckelord
amyloid beta, computer simulation, electrophoresis, electrospray ionization, isoelectric focusing, isotachophoresis, MALDI, mass spectrometry, microchannel, microchip, nano-electrospray ionization, preconcentration, separation
Nationell ämneskategori
Analytisk kemi
Forskningsämne
Kemi
Identifikatorer
urn:nbn:se:kth:diva-193134 (URN)978-91-7729-142-8 (ISBN)
Disputation
2016-11-04, F3, Lindstedtsvägen 26, 10:00 (Engelska)
Opponent
Handledare
Anmärkning

QC 20160930

Tillgänglig från: 2016-09-30 Skapad: 2016-09-29 Senast uppdaterad: 2016-09-30Bibliografiskt granskad

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Jacksén, JohanEmmer, Åsa

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