Titre
High resolution proteome analysis of cryoglobulins using Fourier transform-ion cyclotron resonance mass spectrometry
Type
article
Institution
UNIL/CHUV/Unisanté + institutions partenaires
Périodique
Auteur(s)
Damoc, E.
Auteure/Auteur
Youhnovski, N.
Auteure/Auteur
Crettaz, D.
Auteure/Auteur
Tissot, J. D.
Auteure/Auteur
Przybylski, M.
Auteure/Auteur
Liens vers les personnes
Liens vers les unités
ISSN
1615-9853
Statut éditorial
Publié
Date de publication
2003-08
Volume
3
Numéro
8
Première page
1425
Dernière page/numéro d’article
33
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Aug
Research Support, Non-U.S. Gov't --- Old month value: Aug
Résumé
Cryoglobulins are cold-precipitable serum immunoglobulins associated with a number of infectious, autoimmune and neoplastic disorders such as hepatitis C, Waldenstrom's macroglobulinemia, multiple myeloma, chronic lymphocytic leukemia, and rheumatoid arthritis. The mechanism(s) of cryoprecipitation has remained obscure hitherto, which has prompted recent intensive efforts on the identification of cryoglobulin components. In the present study, two-dimensional gel electrophoresis (2-DE) combined with high resolution Fourier transform-ion cyclotron resonance (FT-ICR) mass spectrometry has been applied as a powerful approach for the analysis of cryoglobulins. While FT-ICR mass spectrometry has been shown to enable the high resolution identification and structure analysis of biopolymers using both electrospray (ESI) and matrix-assisted laser desorption ionization (MALDI), the recently developed MALDI-FT-ICR source is shown here to provide high (sub-ppm) mass determination accuracy and isotopic fine structure as particular advantages in the identification of proteins. The main protein components in a serum cryoprecipitate from a patient with hepatitis C virus (HCV) infection and presenting type II cryogobulinemia are immunoglobulin (Ig)M and IgG which were identified by MALDI-FT-ICR MS analysis after separation by 2-DE as mu- and gamma-heavy chains, kappa- and lambda-light chains, and J-chains. Furthermore, complementarity determining regions CDR1 and CDR2 from monoclonal IgM-RF variable region (V)L were directly identified using accurate mass determinations by FT-ICR-MS. The presence of Spalpha was ascertained as an IgM-associated protein in the serum cryoprecipitate from a patient with HCV infection.
Sujets
PID Serval
serval:BIB_1F6B369576EF
PMID
Date de création
2008-01-25T14:34:33.536Z
Date de création dans IRIS
2025-05-20T15:48:06Z