Rapid identification of hemoglobin variants by electrospray ionization mass spectrometry

被引:88
作者
Wild, BJ
Green, BN
Cooper, EK
Lalloz, MRA
Erten, S
Stephens, AD
Layton, DM
机构
[1] Kings Coll Hosp London, Dept Haematol Med, London SE5 9RS, England
[2] Guys Kings & St Thomas Sch Med, Dept Haematol Med, London SE5 9RS, England
[3] Micromass UK Ltd, Altrincham WA14 5RZ, Cheshire, England
关键词
human; hemoglobin; variant; mass spectrometry; peptide sequence;
D O I
10.1006/bcmd.2001.0430
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
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 mul) but in most cases 2 mul 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. (C) 2001 Academic Press.
引用
收藏
页码:691 / 704
页数:14
相关论文
共 31 条
  • [1] [Anonymous], 1978, BRIT J HAEMATOL, V38, P573
  • [2] BAIN B, 1994, J CLIN PATHOL, V47, P289
  • [3] The laboratory diagnosis of haemoglobinopathies
    Bain, BJ
    Amos, RJ
    Bareford, D
    Chapman, C
    Davies, SC
    Old, JM
    Wild, BJ
    [J]. BRITISH JOURNAL OF HAEMATOLOGY, 1998, 101 (04) : 783 - 792
  • [4] FAST ATOM BOMBARDMENT OF SOLIDS (FAB) - A NEW ION-SOURCE FOR MASS-SPECTROMETRY
    BARBER, M
    BORDOLI, RS
    SEDGWICK, RD
    TYLER, AN
    [J]. JOURNAL OF THE CHEMICAL SOCIETY-CHEMICAL COMMUNICATIONS, 1981, (07) : 325 - 327
  • [5] BHAVNANI M, 1994, J CLIN PATHOL, V47, P199
  • [6] Bissé E, 1998, CLIN CHEM, V44, P2172
  • [7] International Hemoglobin Information Center variant list
    Carver, MFH
    Huisman, THJ
    [J]. HEMOGLOBIN, 1997, 21 (06) : 505 - 506
  • [8] Covey T R, 1988, Rapid Commun Mass Spectrom, V2, P249, DOI 10.1002/rcm.1290021111
  • [9] DECATERINA M, 1992, CLIN CHEM, V38, P1444
  • [10] TANDEM MASS-SPECTROMETRY IN THE CLINICAL ANALYSIS OF VARIANT HEMOGLOBINS
    FALICK, AM
    SHACKLETON, CHL
    GREEN, BN
    WITKOWSKA, HE
    [J]. RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 1990, 4 (10) : 396 - 400