Abbrevations t – fluorescence lifetime j – rotational correlational time ([Ru(bpy)2(phen-ITC)]2+) – Bis-(bipyridine)-5-(isotiocyanatophenantrolin)-Ru(PF6-)2 Ab – antibody AF – Alexa Fluor 660 AF-hGH – with Alexa Fluor 660 labelled human growth hormone FITC – fluorescein isothiocyanate FITC-Ab – FITC labelled antibody FP – fluorescence polarization FRET – fluorescence resonance energy transfer hGH – human growth hormone PMI – N-(3-pyrene)maleimide PMI-hGH – human growth hormone labelled N-(3-pyrene)maleimide PVPPB – poly(N-vinyl-2-pyrolidone) with phenylboronic acid copolymer r – anisotropy RITC – rodamine B isothiocyanate RITC-hGH – with RITC labeled human growth hormone SDS-PAGE – sodium dodecylsulfate-polyacrylamide gel electrophoresis INTRODUCTION Relevance of the study. Human growth hormone (hGH) is a heterogeneous protein with several molecular forms (isoforms). Human growth hormone measurements are complicated because of the heterogeneous nature of hGH. Specific assays for each isoform are currently unavailable. Because the different antibodies used in immunoassays bind to a different spectrum of hGH isoforms, hGH concentrations measured by immunoassay are likely to depend on the particular antibody used. Moreover, because the distribution of the different hGH isoforms varies among individuals, the results from different immunoassays cannot be interrelated easily by using a single conversion factor. Fluorescence is by far the most important optical... [to full text]