title: Novel silent albumin variant (191Ala→Thr) detected by TOF MS of whole plasma.
abstract: BACKGROUND
  Serum protein electrophoresis occasionally reveals multiple albumin bands referred to as bis- or alloalbuminaemia, and whilst the condition can be inherited it may also be acquired.
  
  
  METHODS
  We present a new high resolution approach to the investigation of qualitative changes in albumin structure. The on-line reverse phase time-of-flight mass spectrometry procedure (TOF MS) elaborated here requires <0.2 μl of plasma and takes ~10 min to perform. Two plasma samples with classical bisalbuminaemia were used to verify the efficacy of the procedure for detecting genetic variants. When a novel mutation was detected, mass mapping of native unreduced albumin was used to pinpoint its location and inform targeted DNA sequencing of the albumin gene.
  
  
  RESULTS
  Normal serum albumin showed its expected major isoform at 66,439 Da and the electrophoretic variants showed co-equal expression of additional components at -14 and +744 Da respectively. Surprisingly, one of the supposed controls showed paired albumin peaks at 66,439 and 66,469 Da and this 30 Da increase in mass was localised to between Arg(114) and Arg(197) and confirmed as being due to a novel 191Ala→Thr (+30 Da) substitution through DNA sequencing.
  
  
  CONCLUSIONS
  The electrospray TOF MS approach developed here provides a rapid, sensitive and extremely precise method of revealing minute changes in albumin primary structure.
authors.affiliation: Pathology Department, University of Otago, Christchurch, New Zealand; Canterbury Health Laboratories, Christchurch, New Zealand. Electronic address: steve.brennan@otago.ac.nz.
authors.forename: Stephen O
authors.initials: SO
authors.lastname: Brennan
authors.affiliation: Canterbury Health Laboratories, Christchurch, New Zealand.
authors.forename: Catherine
authors.initials: C
authors.lastname: Rollo
authors.affiliation: Canterbury Health Laboratories, Christchurch, New Zealand.
authors.forename: Howard C
authors.initials: HC
authors.lastname: Potter
mesh_terms: D019943:Amino Acid Substitution; D006505:Hepatitis / Q000097:blood; D006801:Humans; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D017422:Sequence Analysis, DNA; D012709:Serum Albumin / Q000032:analysis* / Q000737:chemistry / Q000235:genetics*
chemical_list: D012709:Serum Albumin
keywords: Amino acid substitution; Blood proteins; Post translational modification; Serum albumin; Whole protein mass spectrometry