Immunalysis Corporation, 829 Towne Center Drive, Pomona, CA 91767, USA.
Drug Test Anal. 2009 Sep;1(9-10):441-6. doi: 10.1002/dta.101.
Approaches to detect whether an athlete has used growth hormone have been intensely investigated by sport organizations for 20 years. This effort has led to a human growth hormone (hGH) isoform ratio test in serum that has been approved by WADA and deployed at three Olympic Games, although a positive case has yet to be reported. We set out to determine whether the ratio test could be applied to urine. First we investigated various ways to extract hGH from spiked urine. We were able to recover 95% using selective centrifugal concentration. This fraction was then subjected to four different commercially available immunoprecipitation kits. The highest yield was obtained with the Invitrogen Dynabeads Protein G kit. Nevertheless it is apparent that these methods do not recover enough hGH for subsequent analysis by mass spectrometry. With further effort greater recovery of the 22 kDa isoform might be achieved, however it is very unlikely that the 20 kDa isoform could be detected. This method may be significantly improved by the application of both nanoparticle and aptamer technology.
二十年来，体育组织一直在深入研究检测运动员是否使用生长激素的方法。这一努力促使了血清中人类生长激素（hGH）同工型比例测试的出现，该测试已被世界反兴奋剂机构（WADA）批准，并在三届奥运会上使用，尽管尚未报告阳性病例。我们着手确定该比例测试是否可应用于尿液。首先，我们研究了从加标尿液中提取 hGH 的各种方法。我们使用选择性离心浓缩能够回收 95%。然后，将该馏分进行四种不同的市售免疫沉淀试剂盒的测试。使用 Invitrogen Dynabeads Protein G 试剂盒获得的产量最高。然而，很明显，这些方法无法回收足够的 hGH 用于随后的质谱分析。通过进一步的努力，可能会实现对 22 kDa 同工型的更高回收率，但几乎不可能检测到 20 kDa 同工型。通过纳米颗粒和适体技术的应用，该方法可能会得到显著改进。
