Bioanalysis Group, Neuroscience Research Program, IMIM-Parc Salut Mar, Barcelona Biomedical Research Park, Barcelona, Spain.
Anal Chim Acta. 2012 Jul 6;733:56-63. doi: 10.1016/j.aca.2012.04.028. Epub 2012 May 3.
Detecting recombinant human growth hormone (rhGH) abuse in sport remains one of the major challenges in doping control. We have compared two different approaches to detect the hGH (human growth hormone) abuse. The first measures the concentrations of the 22 kDa hGH isoform (rec assay) and pituitary derived isoforms (pit assay) and a ratio rec/pit is obtained. The second measures the concentrations of 22 and 20 kDa hGH isoforms and also a ratio 22/20 kDa is derived. Using a single set (nine healthy male subjects, 7 days, 0.026 mg/kg/day of rhGH, 2 week wash out period) both approaches were compared. To quantify the agreement between the immunoassays, B.A. (Bland-Altman) analysis and P.r. (Pearson correlation) were used. To fully understand the assay readings, all relevant antibodies were characterised by surface plasmon resonance (SPR). In either approach the ratio numerator produces similar results and the denominator determines both signal-amplitude and time-frame of possible application. The rec vs pit approach displays a higher distinctive capacity to detect hGH abuse but the complex binding properties of the capture antibodies make it very difficult to evaluate the precise contributions of the individual hGH variants to the assay result. In the 22 vs 20 approach, the 20 kDa hGH concentration measures determine its applicability. Both approaches are based on a different principle, should be preferably applied within 24 h after rhGH administration, and are perfectly comparable given the results obtained. The reduced time frame of application indicates that their principle application should be preferably in an out-of-competition setting.
检测运动中重组人生长激素 (rhGH) 的滥用仍然是兴奋剂控制中的主要挑战之一。我们比较了两种不同的方法来检测 hGH(人生长激素)的滥用。第一种方法测量 22 kDa hGH 同工型(rec 测定)和垂体衍生同工型(pit 测定)的浓度,并获得 rec/pit 比值。第二种方法测量 22 和 20 kDa hGH 同工型的浓度,也得出 22/20 kDa 的比值。使用一组(九名健康男性受试者,7 天,0.026 mg/kg/天 rhGH,2 周洗脱期)比较两种方法。为了定量评估免疫测定之间的一致性,使用 B.A.(Bland-Altman)分析和 P.r.(Pearson 相关)。为了全面理解测定结果,所有相关抗体均通过表面等离子体共振 (SPR) 进行了表征。在这两种方法中,比值的分子产生相似的结果,而分母决定了信号幅度和可能应用的时间框架。rec 与 pit 的方法显示出更高的区分能力来检测 hGH 滥用,但捕获抗体的复杂结合特性使得很难评估个体 hGH 变体对测定结果的精确贡献。在 22 与 20 的方法中,20 kDa hGH 浓度的测量决定了其适用性。这两种方法基于不同的原理,最好在 rhGH 给药后 24 小时内应用,并且考虑到所获得的结果,它们是完全可比的。应用时间框架的缩短表明,其原理应用最好是在非比赛环境中。