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Molecularly imprinted polymers coupled to matrix assisted laser desorption ionization mass spectrometry for femtomoles detection of cardiac troponin I peptides
Author(s) -
Cenci Lucia,
Anesi Andrea,
Busato Mirko,
Guella Graziano,
Bossi Alessandra Maria
Publication year - 2016
Publication title -
journal of molecular recognition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.401
H-Index - 79
eISSN - 1099-1352
pISSN - 0952-3499
DOI - 10.1002/jmr.2494
Subject(s) - chemistry , chromatography , molecularly imprinted polymer , matrix assisted laser desorption/ionization , mass spectrometry , peptide , troponin i , detection limit , desorption , polymer , selectivity , adsorption , biochemistry , organic chemistry , psychology , psychiatry , myocardial infarction , catalysis
Molecularly imprinted polymers (MIPs) were combined to MALDI‐TOF‐MS to evaluate a selective enrichment (SE) method for the determination of clinically relevant biomarkers from complex biological samples. The concept was proven with the myocardial injury marker Troponin I (cTnI). In a first part, MIP materials entailed for the recognition of cTnI epitopes (three peptides selected) were prepared and characterized in dimensions (0.7–2μm), dissociation constants (58–817 nM), kinetics of binding (5–60 min), binding capacity (ca. 1.5 µg/mg polymer), imprinting factors (3 > IF > 5) and selectivity for the peptide epitope. Then, the MIPs, incubated with cTnI peptides and spotted on the target with the DHB matrix, were assayed for the desorption of the peptides in MALDI‐TOF‐MS. The measured detection limit was ca. 300 femtomols. Finally, the MIP‐SE MALDI‐TOF‐MS was tested for its ability to enrich in the cTnI peptides from a complex sample, mimic of serum (i.e. 81 peptides of digested albumin). The MIP‐SE MALDI‐TOF‐MS successfully enriched in cTnI peptides from the complex sample proving the technique could offer a flexible platform to prepare entailed materials suitable for diagnostic purposes. Copyright © 2015 John Wiley & Sons, Ltd.