title: Rapid identification of hemoglobin variants by electrospray ionization mass spectrometry.
abstract: The precise identification of human hemoglobin variants, over 700 human hemoglobin variants are known, is essential for prediction of their clinical and genetic significance. A systematic approach to their rapid identification is described. Traditionally this requires protein or DNA characterization which entails lengthy analytical procedures. To overcome these obstacles a rapid approach to variant hemoglobin identification has been developed using conventional phenotypic methods combined with electrospray ionization-mass spectrometry (ESI-MS). The latter requires only a small amount of whole blood (10 microl) but in most cases 2 microl would have been sufficient and no preanalytical steps, such as separation of red cells or globin chains, are necessary. Aged, hemolyzed blood samples can also be analyzed. This approach has been used to positively identify 95% of the variants in over 250 samples. The remaining 5% in which a variant was detected by phenotypic techniques were not resolved by mass spectrometry. Ninety-nine different abnormalities comprising 36 alpha-chain variants, 59 beta-chain variants (including 2 extensions), and 4 hybrid hemoglobins were identified. These include 15 novel variants. The application of ESI-MS described requires approximately 1 h to prepare and analyze each sample and has minimal reagent costs. The turnaround time on a single sample can be as little as 2 h. This technique can now be considered a useful additional tool for reference laboratories.
authors.affiliation: Department of Haematological Medicine, Guy's, King's, and St. Thomas' School of Medicine, Denmark Hill Campus, London SE5 9RS, UK. barbara.wild@kcl.ac.uk
authors.forename: B J
authors.initials: BJ
authors.lastname: Wild
authors.forename: B N
authors.initials: BN
authors.lastname: Green
authors.forename: E K
authors.initials: EK
authors.lastname: Cooper
authors.forename: M R
authors.initials: MR
authors.lastname: Lalloz
authors.forename: S
authors.initials: S
authors.lastname: Erten
authors.forename: A D
authors.initials: AD
authors.lastname: Stephens
authors.forename: D M
authors.initials: DM
authors.lastname: Layton
mesh_terms: D019943:Amino Acid Substitution; D004252:DNA Mutational Analysis; D014644:Genetic Variation / Q000235:genetics*; D005914:Globins / Q000235:genetics; D006455:Hemoglobins, Abnormal / Q000032:analysis* / Q000235:genetics; D006801:Humans; D008722:Methods; D008832:Microchemistry; D013048:Specimen Handling; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods* / Q000592:standards
chemical_list: D006455:Hemoglobins, Abnormal; D005914:Globins