title: Quantitative mass spectrometry analysis of intact hemoglobin A2 by precursor ion isolation and detection.
abstract: Precise and accurate quantification of proteins is essential in clinical laboratories. Here, we present a mass spectrometry (MS)-based method for the quantification of intact proteins in an ion trap mass spectrometer. The developed method is based on the isolation and detection of precursor ions for the quantification of the corresponding signals. The method was applied for the quantification of hemoglobin (Hb) A2, a marker used for the diagnosis of a β-thalassemia trait. The α and δ globin chains, corresponding to total Hb and HbA2, respectively, were isolated in the ion trap at specific charge states and ejected without activation. Areas of the corresponding isolated precursor ions were used to calculate the δ to α ratio. Three series of quantifications were performed on 7 different days. The standard curve fitted linearly (R(2) = 0.9982) and allowed quantification of HbA2 over a concentration range from 3% to 18% of total Hb. Analytical imprecision ranged from 3.5% to 5.3%, which is enough to determine if the HbA2 level is below 3.5% or above 3.7%. In conclusion, our method reaches precision requirements that would be acceptable for the quantitative measurement of diagnostic proteins, such as HbA2, in clinical laboratories.
authors.affiliation: Department of Human Protein Sciences, Faculty of Medicine, Geneva University, Geneva, Switzerland.
authors.forename: Adelina E
authors.initials: AE
authors.lastname: Acosta-Martin
authors.forename: Didia
authors.initials: D
authors.lastname: Coelho Graça
authors.forename: Paola
authors.initials: P
authors.lastname: Antinori
authors.forename: Lorella
authors.initials: L
authors.lastname: Clerici
authors.forename: Ralf
authors.initials: R
authors.lastname: Hartmer
authors.forename: Markus
authors.initials: M
authors.lastname: Meyer
authors.forename: Denis
authors.initials: D
authors.lastname: Hochstrasser
authors.forename: Kaveh
authors.initials: K
authors.lastname: Samii
authors.forename: Pierre
authors.initials: P
authors.lastname: Lescuyer
authors.forename: Alexander
authors.initials: A
authors.lastname: Scherl
mesh_terms: D015415:Biomarkers / Q000032:analysis*; D006443:Hemoglobin A2 / Q000032:analysis*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D017086:beta-Thalassemia / Q000175:diagnosis
chemical_list: D015415:Biomarkers; D006443:Hemoglobin A2