Ribela M T, Bianco A C, Bartolini P
Department of Application of Nuclear Techniques in Biological Sciences, University of São Paulo, Brazil.
J Clin Endocrinol Metab. 1996 Jan;81(1):249-56. doi: 10.1210/jcem.81.1.8550760.
Recombinant human TSH (rec-hTSH; Thyrogen, lot M-17073) obtained from transformed Chinese hamster ovary cells was tested for both radioiodination and preparation of a secondary standard used in RIA and immunoradiometric assay (IRMA) for routine clinical investigation. Results were compared to those obtained with high quality pituitary TSH (pit-hTSH; Dr. P. Torjesen, Oslo, Norway; and NIDDK, Rockville, MD), traditionally used in these assays. After extensive characterization and testing, it was found that [125I]rec-hTSH matched all binding and chromatographic criteria usually obtained with [125I]pit-hTSH, including Stokes' radius, labeling, and storage stability, and did not introduce any significant bias when used in the measurement of unknown serum samples. A preparation of rec-hTSH was calibrated against a local secondary standard as well as against two well known international reference preparations (NIDDK hTSH RP-1 and WHO International Reference Preparation 80/558) by IRMA and RIA. In the RIA, NIDDK anti-hTSH-3 polyclonal antibody was used, whereas in the IRMA, two commercial preparations were used: a monoclonal antibody as the detecting antibody, and a polyclonal antibody as the capture antibody. In both assays, the recombinant standard preparation yielded good fit displacement curves, showing significant parallelism compared to pit-hTSH and therefore allowing an unbiased measurement of unknown serum samples. The specific activity of the rec-hTSH preparation calibrated against the WHO International Reference Preparation was 7.7 IU/mg protein when measured by IRMA and 7.1 IU/mg when measured by RIA. In conclusion, these results indicate for the first time that rec-hTSH can fully replace pit-hTSH as both standard and tracer in diagnostic in vitro systems such as RIA and IRMA, suggesting that other recombinant glycosylated hormones might also serve for immunoassay reagent preparation.
对从转化的中国仓鼠卵巢细胞中获得的重组人促甲状腺激素（rec-hTSH；Thyrogen，批次M-17073）进行了放射性碘化测试，并制备了用于放射免疫分析（RIA）和免疫放射分析（IRMA）的二级标准品，用于常规临床研究。将结果与使用传统上用于这些分析的高质量垂体促甲状腺激素（pit-hTSH；挪威奥斯陆的P. Torjesen博士和美国马里兰州罗克维尔的美国国立糖尿病、消化和肾脏疾病研究所提供）所获得的结果进行比较。经过广泛的表征和测试，发现[125I]rec-hTSH符合通常用[125I]pit-hTSH获得的所有结合和色谱标准，包括斯托克斯半径、标记和储存稳定性，并且在用于测量未知血清样本时不会引入任何显著偏差。通过IRMA和RIA，根据当地二级标准品以及两种知名的国际参考制剂（美国国立糖尿病、消化和肾脏疾病研究所hTSH RP-1和世界卫生组织国际参考制剂80/558）对rec-hTSH制剂进行校准。在RIA中，使用美国国立糖尿病、消化和肾脏疾病研究所抗hTSH-3多克隆抗体，而在IRMA中，使用两种商业制剂：一种单克隆抗体作为检测抗体，一种多克隆抗体作为捕获抗体。在这两种分析中，重组标准制剂均产生了良好的拟合置换曲线，与pit-hTSH相比显示出显著的平行性，因此能够对未知血清样本进行无偏差测量。根据世界卫生组织国际参考制剂校准的rec-hTSH制剂的比活性，通过IRMA测量时为7.7 IU/mg蛋白质，通过RIA测量时为7.1 IU/mg。总之，这些结果首次表明rec-hTSH可以在RIA和IRMA等体外诊断系统中完全替代pit-hTSH作为标准品和示踪剂，这表明其他重组糖基化激素也可能用于免疫分析试剂的制备。
