title: Detection of recombinant growth hormone in human plasma by a 2-D PAGE method.
abstract: Human growth hormone (GH) has several central metabolic functions including bone growth in childhood, and its anabolic and lipolytic effects in particular are assumed reasons for the abuse of GH by athletes. Human endogenous GH consists of a main 22 kDa variant and several isoforms. In contrast, recombinant GH consists of only one variant being identical to the main endogenous isoform. The method presented here separates different isoforms by 2-D PAGE after isolation of GH from plasma using an immunoaffinity purification system. While samples containing endogenous GH yield up to four isoforms, samples with recombinant GH contain the main 22 kDa spot only. Normalized spot volumes (NSV) are calculated after addition of an internal standard and a discrimination limit was determined at 0.52 for the NSV of the main 22 kDa spot. Above this value, samples containing endogenous GH show at least the main 22 kDa isoform and the 20 kDa splice variant. In contrast, samples with a NSV >0.52 and only one spot are suspicious to contain recombinant GH. This method detects discrete isoforms of GH from plasma and discriminates endogenous GH from its recombinant analog, which makes it useful for doping control purposes.
authors.affiliation: Institute of Biochemistry/Center for Preventive Doping Research, German Sport University Cologne, Germany.
authors.forename: Maxie
authors.initials: M
authors.lastname: Kohler
authors.forename: Klaus
authors.initials: K
authors.lastname: Püschel
authors.forename: Dimitri
authors.initials: D
authors.lastname: Sakharov
authors.forename: Alexander
authors.initials: A
authors.lastname: Tonevitskiy
authors.forename: Wilhelm
authors.initials: W
authors.lastname: Schänzer
authors.forename: Mario
authors.initials: M
authors.lastname: Thevis
mesh_terms: D000328:Adult; D004300:Doping in Sports; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005260:Female; D019382:Human Growth Hormone / Q000097:blood* / Q000378:metabolism; D006801:Humans; D016014:Linear Models; D008297:Male; D010902:Pituitary Gland / Q000737:chemistry; D011994:Recombinant Proteins / Q000097:blood; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D011994:Recombinant Proteins; D019382:Human Growth Hormone