dc.contributor |
Universitat de Barcelona |
dc.contributor.author |
Peró Gascón, Roger |
dc.contributor.author |
Pont Villanueva, Laura |
dc.contributor.author |
Benavente Moreno, Fernando J. (Julián) |
dc.contributor.author |
Barbosa Torralbo, José |
dc.contributor.author |
Sanz Nebot, María Victoria |
dc.date |
2018-06-21T14:04:35Z |
dc.date |
2018-06-21T14:04:35Z |
dc.date |
2016 |
dc.date |
2018-06-21T14:04:36Z |
dc.identifier.citation |
0173-0835 |
dc.identifier.citation |
655669 |
dc.identifier.uri |
http://hdl.handle.net/2445/123187 |
dc.format |
12 p. |
dc.format |
application/pdf |
dc.language.iso |
eng |
dc.publisher |
Wiley-VCH |
dc.relation |
Versió postprint del document publicat a: https://doi.org/10.1002/elps.201500495 |
dc.relation |
Electrophoresis, 2016, vol. 37, p. 1220-1231 |
dc.relation |
https://doi.org/10.1002/elps.201500495 |
dc.rights |
(c) Wiley-VCH, 2016 |
dc.rights |
info:eu-repo/semantics/openAccess |
dc.subject |
Electroforesi capil·lar |
dc.subject |
Espectrometria de masses |
dc.subject |
Camps magnètics |
dc.subject |
Capillary electrophoresis |
dc.subject |
Mass spectrometry |
dc.subject |
Magnetic fields |
dc.title |
Analysis of serum transthyretin by on-line immunoaffinity solid-phase extraction capillary electrophoresis mass spectrometry using magnetic beads |
dc.type |
info:eu-repo/semantics/article |
dc.type |
info:eu-repo/semantics/acceptedVersion |
dc.description.abstract |
In this paper, an on-line immunoaffinity solid-phase extraction capillary electrophoresis mass spectrometry (IA-SPE-CE-MS) method using magnetic beads (MBs) is described for the analysis of serum transthyretin (TTR), which is a protein related to different types of amyloidosis. First, purification of TTR from serum was investigated by off-line immunoprecipitation and CE-MS. The suitability of three Protein A (ProA) MBs (Protein A Ultrarapid AgaroseTM (UAPA), Dynabeads® Protein A (DyPA) and SiMAG-Protein A (SiPA)) and AffiAmino Ultrarapid AgaroseTM (UAAF) MBs to prepare an IA sorbent with a polyclonal antibody (Ab) against TTR, was studied. In all cases results were repeatable and it was possible the identification and the quantitation of the relative abundance of the 6 most abundant TTR proteoforms. Although recoveries were the best with UAPA MBs, UAAF MBs were preferred for on-line immunopurification because Ab was not eluted from the MBs. Under the optimised conditions with standards in IA-SPE-CE-MS, microcartridge lifetime (>20 analyses/day) and repeatability (2.9 and 4.3 % RSD for migration times and peak areas) were good, the method was linear between 5- 25 µg·mL-1 and limit of detection (LOD) was around 1 µg·mL-1 (25 times lower than by CE-MS, 25 µg·mL-1). A simple off-line sample pretreatment based on precipitation of the most abundant proteins with 5% (v/v) of phenol was necessary to clean-up serum samples. The potential of the on-line method to screen for familial amyloidotic polyneuropathy type I (FAP-I), which is the most common hereditary systemic amyloidosis, was demonstrated analysing serum samples from healthy controls and FAP-I patients. |