title: Plasma myostatin measured by a competitive ELISA using a highly specific antiserum.
abstract: BACKGROUND
  Understanding the (patho)physiology of the negative muscle regulator myostatin (Myo) is important for patients with skeletal muscle disorders or cardiac disease. However, a reliable tool for measuring plasma Myo immunoreactivity is still lacking.
  
  
  METHODS
  Human full-length proMyo was used to raise a polyclonal rabbit antiserum for a competitive Myo ELISA that was validated in patients with decompensated congestive heart failure (CHF) and in control patients (n=20 each).
  
  
  RESULTS
  The Myo antiserum detected all subunits of human proMyo. The calibration curve showed an optimal range between 0.3 and 83.3 ng/ml (7.5-2100 pmol/l), with no cross-reactivity to growth differentiation factor-11, follistatin and follistatin-related gene protein. The inter-assay and intra-assay variances in human serum were ≤15% and ≤10%, respectively; the detection limit was 270 pg/ml (6.75 pmol/l). The assay showed excellent linearity in human plasma. Plasma NT-proBNP and Myo were significantly elevated in decompensated CHF compared with control patients and decreased significantly upon recompensating therapy.
  
  
  CONCLUSIONS
  We describe the development of the first ELISA for myostatin immunoreactivity and its validation during recompensating therapy for CHF. This assay will be valuable for investigating neurological and cardiac diseases and states of cachexia, insulin resistance, and obesity.
authors.affiliation: Immundiagnostik AG, Stubenwald-Allee 8a, 64625 Bensheim, Germany.
authors.forename: Karl Florian
authors.initials: KF
authors.lastname: Wintgens
authors.forename: Thomas
authors.initials: T
authors.lastname: Dschietzig
authors.forename: Stanka
authors.initials: S
authors.lastname: Stoeva
authors.forename: Mats
authors.initials: M
authors.lastname: Paulsson
authors.forename: Franz Paul
authors.initials: FP
authors.lastname: Armbruster
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D016022:Case-Control Studies; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006333:Heart Failure / Q000097:blood* / Q000175:diagnosis; D006801:Humans; D007106:Immune Sera; D008297:Male; D008875:Middle Aged; D055435:Myostatin / Q000097:blood* / Q000235:genetics / Q000276:immunology; D020097:Natriuretic Peptide, Brain / Q000097:blood; D010446:Peptide Fragments / Q000097:blood; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D007106:Immune Sera; C525911:MSTN protein, human; D055435:Myostatin; D010446:Peptide Fragments; D011994:Recombinant Proteins; D020097:Natriuretic Peptide, Brain