Detection of recombinant growth hormone in human plasma by a 2-D PAGE method.
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Detection of recombinant growth hormone in human plasma by a 2-D PAGE method. / Kohler, Maxie; Püschel, Klaus; Sakharov, Dimitri; Tonevitskiy, Alexander; Schänzer, Wilhelm; Thevis, Mario.
In: ELECTROPHORESIS, Vol. 29, No. 22, 22, 2008, p. 4495-4502.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Detection of recombinant growth hormone in human plasma by a 2-D PAGE method.
AU - Kohler, Maxie
AU - Püschel, Klaus
AU - Sakharov, Dimitri
AU - Tonevitskiy, Alexander
AU - Schänzer, Wilhelm
AU - Thevis, Mario
PY - 2008
Y1 - 2008
N2 - 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.
AB - 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.
M3 - SCORING: Zeitschriftenaufsatz
VL - 29
SP - 4495
EP - 4502
JO - ELECTROPHORESIS
JF - ELECTROPHORESIS
SN - 0173-0835
IS - 22
M1 - 22
ER -