title: Detection of transthyretin variants using immunoprecipitation and matrix-assisted laser desorption/ionization bioreactive probes: a clinical application of mass spectrometry.
abstract: In our continuing efforts to develop mass spectrometry-based methods for transthyretin (TTR) variant detection and characterization, we have sought to use matrix-assisted laser desorption/ionization (MALDI) bioreactive probes incorporating immobilized trypsin for screening purposes. These devices show good diagnostic potential as a clinical screening tool to detect amino acid substitutions in TTR. MALDI probes allow the on-probe generation of tryptic digests. The subsequent mass analysis of the on-probe digest yields the peptide map. The inherent advantages of this method include considerably reduced digestion times (minutes vs. hours), absence of autolysis products, minimized sample handling, and hence minimal sample loss. A further advantage is that the opportunity for loss of hydrophobic peptides is reduced because no sample transfer occurs. The method can be applied as a preliminary screen for TTR variants where TTR is isolated from patient serum through immunoprecipitation. This method should also be applicable to other proteins and suitable for automation.
authors.affiliation: Mass Spectrometry Resource, Boston University School of Medicine, Massachusetts 02118, USA.
authors.forename: R
authors.initials: R
authors.lastname: Théberge
authors.forename: L H
authors.initials: LH
authors.lastname: Connors
authors.forename: M
authors.initials: M
authors.lastname: Skinner
authors.forename: C E
authors.initials: CE
authors.lastname: Costello
mesh_terms: D006454:Hemoglobins / Q000737:chemistry; D006801:Humans; D006868:Hydrolysis; D007202:Indicators and Reagents; D008970:Molecular Weight; D011228:Prealbumin / Q000737:chemistry*; D011233:Precipitin Tests; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D006454:Hemoglobins; D007202:Indicators and Reagents; D011228:Prealbumin