Mass spectrometric analysis of human transferrin in different body fluids.
In this study, we present a versatile new procedure for the analysis of transferrin and its isoforms isolated from human body fluids such as serum, plasma, and cerebrospinal fluid. This method is based on a three-step procedure: (i) isolation of transferrins using anion-exchange chromatography with UV detection; (ii) concentration of the transferrin fraction; (iii) detection of the transferrins with liquid chromatography-electrospray mass spectrometry. Pre-analytical sample procedures can be omitted and no immunoaffinity columns or transferrin-specific immunoassays were used. Anticoagulants such as heparin, EDTA, citrate, and oxalate do not interfere with our analysis. According to their respective molecular masses, up to ten different isoforms of transferrin could be identified in a serum sample from a patient with a congenital disorder of glycosylation type Ia (CDG-Ia). The method was successfully applied to different pathological samples from patients with CDG-Ia, CDG-Ib, CDG-Ic, CDG-Ie, CDG-If, and CDG-IIa. Additionally, samples from alcohol consumers that were found with turbidimetric immunoassay to contain increased levels of carbohydrate-deficient transferrin were analyzed.
Kleinert
Peter
P
Division of Clinical Chemistry and Biochemistry, Department of Pediatrics, University of Zurich, Zurich, Switzerland.
Kuster
Thomas
T
Durka
Silke
S
Ballhausen
Diana
D
Bosshard
Nils U
NU
Steinmann
Beat
B
Hänseler
Edgar
E
Jaeken
Jaak
J
Heizmann
Claus W
CW
Troxler
Heinz
H
D001826:Body Fluids / Q000737:chemistry*; D002239:Carbohydrate Metabolism, Inborn Errors / Q000175:diagnosis; D002853:Chromatography, Liquid; D006801:Humans; D020033:Protein Isoforms / Q000032:analysis; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D014168:Transferrin / Q000032:analysis* / Q000302:isolation & purification
D020033:Protein Isoforms; D014168:Transferrin