title: Derivatization strategies for CE-LIF analysis of biomarkers: Toward a clinical diagnostic of familial transthyretin amyloidosis.
abstract: We report three derivatization strategies for CE analysis with LIF detection (CE-LIF) of two synthetic peptides mimicking the wild and mutated fragments of interest for the diagnosis of familial transthyretin amyloidosis. The precapillary derivatization of the peptides with three optical tags, 5-carboxytetramethylrhodamin succinimidyl ester (TAMRA-SE), naphtalene-2,3-dicarboxyaldehyde (NDA), and 3-(2-furoyl)quinoline-2-carboxyaldehyde (FQ) has been investigated by CE-LIF detection and MS. Results provide evidence that high reaction yields have been reached whereas the multitagging phenomenon has occurred for both NDA and TAMRA-SE labeling procedures. The derivatization and electrokinetic separation of a mixture of the two peptides of interest for the pathology diagnosis (22-aa peptides that differ only from one amino acid) were achieved using both approaches. The highest resolution with a value of 2.5 was obtained with TAMRA-SE labeled derivatives whereas NDA gave the best detection sensitivity (LOD of 2.5 μM). The validation of the developed methods showed a good linearity (R ≥ 0.997) between the peak area of the labeled derivatives and the peptide concentration for both NDA and FQ labeling procedures. The intraday RSDs of A and the migration times were less than 3.8 and 2.2%, respectively.
authors.affiliation: Faculté de Pharmacie, Université Paris-Sud, Châtenay-Malabry, France; CNRS, Institut Galien Paris-Sud, UMR 8612, Châtenay-Malabry, France.
authors.forename: Sonia
authors.initials: S
authors.lastname: Korchane
authors.forename: Antoine
authors.initials: A
authors.lastname: Pallandre
authors.forename: Cédric
authors.initials: C
authors.lastname: Przybylski
authors.forename: Christian
authors.initials: C
authors.lastname: Poüs
authors.forename: Florence
authors.initials: F
authors.lastname: Gonnet
authors.forename: Myriam
authors.initials: M
authors.lastname: Taverna
authors.forename: Régis
authors.initials: R
authors.lastname: Daniel
authors.forename: Isabelle
authors.initials: I
authors.lastname: Le Potier
mesh_terms: D028227:Amyloid Neuropathies, Familial / Q000097:blood / Q000175:diagnosis*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005456:Fluorescent Dyes; D006801:Humans; D016014:Linear Models; D008956:Models, Chemical; D010455:Peptides / Q000032:analysis* / Q000737:chemistry*; D011228:Prealbumin / Q000032:analysis / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes; D010455:Peptides; D011228:Prealbumin
keywords: CE; Familial transthyretin amyloidosis; LIF; MS; Peptide labeling