title: Analysis of urinary human growth hormone (hGH) using hydrogel nanoparticles and isoform differential immunoassays after short recombinant hGH treatment: preliminary results.
abstract: Successful application clinical-grade human growth hormone (hGH) immunoassays to the discovery of illegal doping cases has been rare. Indeed, the preferred biological matrix in doping control is urine, where the estimated baseline concentration of hGH falls well below the linear range and sensitivity threshold of all commercially available immunoassays, including hGH isoform differential immunoassays which can discriminate pituitary endogenous hGH from recombinant hGH. We employed hydrogel nanoparticles as a pre-processing step that concentrate urinary hGH into the linear range of isoform differential immunoassays. We explored the characteristics of immunoassays in urine spiked with both phGH or rhGH, after pre-treatment with the nanoparticles. Subsequently, pre-treatment was applied to urine obtained from 3 healthy volunteers administered during three days with daily subcutaneous injections of 0.026 mg/kg/day rhGH, Genotonorm(®). Linearity between both rhGH and phGH concentrations in urine measured by a chemoluminescent assay (Immulite) and in the particle eluate was evident for differential immunoassays (R square higher than 0.999). In case of treated individuals the recombinant/pituitary concentration ratios remained above the established World Anti-Doping Agency (WADA) criterion for hGH misuse up to 24h after the last administration dose, using both assays for volunteer 1 and 2 while in case of volunteer 3 results were inconclusive. The use of nanoparticles appears to open the possibility of assessing rhGH misuse in urine.
authors: Bioanalysis Group IMIM-Parc Salut Mar and Department of Experimental and Health Sciences, Universitat Pompeu Fabra, Barcelona Biomedical Research Park (PRBB), Barcelona, Spain.
authors: Jaume
authors: J
authors: Bosch
authors: Alessandra
authors: A
authors: Luchini
authors: Simona
authors: S
authors: Pichini
authors: Davide
authors: D
authors: Tamburro
authors: Claudia
authors: C
authors: Fredolini
authors: Lance
authors: L
authors: Liotta
authors: Emanuel
authors: E
authors: Petricoin
authors: Roberta
authors: R
authors: Pacifici
authors: Francesco
authors: F
authors: Facchiano
authors: Jordi
authors: J
authors: Segura
authors: Enrico
authors: E
authors: Garaci
authors: Ricardo
authors: R
authors: Gutiérrez-Gallego
mesh_terms: D019382:Human Growth Hormone / Q000652:urine*; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D007118:Immunoassay / Q000379:methods*; D008297:Male; D053758:Nanoparticles / Q000737:chemistry*; D020033:Protein Isoforms / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D020033:Protein Isoforms; D011994:Recombinant Proteins; D019382:Human Growth Hormone; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate
keywords: AAF; Differential immunoassays; Hydrogel nanoparticles; Pit; Pituitary growth hormone; Rec; Recombinant growth hormone; adverse analytical finding; hGH; human growth hormone; pituitary; recombinant
references: Curr Mol Med. 2010 Mar;10(2):133-41
references: 20196732
references: Endocr Rev. 2012 Apr;33(2):155-86
references: 22368183
references: Anal Chim Acta. 2012 Jul 6;733:56-63
references: 22704376
references: Drug Test Anal. 2009 Sep;1(9-10):447-54
references: 20355230
references: Ther Drug Monit. 2009 Feb;31(1):3-13
references: 19155963
references: Clin Chem. 2009 Mar;55(3):445-53
references: 19168559
references: Nano Lett. 2008 Jan;8(1):350-61
references: 18076201
references: Nano Res. 2008 Dec;1(6):502-518
references: 20467576
references: Am J Physiol Endocrinol Metab. 2002 Oct;283(4):E836-43
references: 12217902
references: J Sports Med Doping Stud. 2012;2(6):
references: 24014257
references: J Mater Chem. 2009 Aug 7;19(29):5071-5077
references: 20585471