Egrie J C, Cotes P M, Lane J, Gaines Das R E, Tam R C
J Immunol Methods. 1987 May 20;99(2):235-41. doi: 10.1016/0022-1759(87)90133-5.
Since the estimation of erythropoietin (EPO) by radioimmunoassay (RIA) has been limited by the availability of highly purified urinary (U) human (Hu) EPO, we investigated the use of recombinant (R)-HuEPO as a replacement for U-HuEPO in the preparation of 125I-tracer and high affinity antisera. In each of two validated RIAs developed using U-HuEPO-derived reagents, dose-response lines and potency estimates for samples were compared when 125I-U-HuEPO and anti-U-HuEPO antisera were sequentially replaced to give assay variants using R-HuEPO-derived reagents. Two U-HuEPO preparations, the 2nd International Reference Preparation and CAT-1, and R-HuEPO were variously used as standards. The samples tested included clinically relevant human sera and partially purified preparations of U-HuEPO and R-HuEPO. In each RIA and for each assay variant tested, samples gave dose-response lines whose slopes did not differ significantly from that of the standard. For each of the two variant RIAs, potency estimates for any sample were consistent and, where examined, RIA potency estimates agreed with in vivo bioassay determinations. These results, obtained independently in two laboratories, indicate that RIAs having appropriate specificity and sensitivity for the estimation of EPO in clinical samples can be developed using reagents derived from R-HuEPO.
由于放射免疫分析（RIA）对促红细胞生成素（EPO）的检测受到高纯度尿源性人促红细胞生成素（U-HuEPO）可得性的限制，我们研究了使用重组人促红细胞生成素（R-HuEPO）替代U-HuEPO来制备¹²⁵I标记物和高亲和力抗血清。在使用U-HuEPO衍生试剂开发的两种经过验证的放射免疫分析中，当¹²⁵I-U-HuEPO和抗U-HuEPO抗血清依次被R-HuEPO衍生试剂替代以获得分析变体时，比较了剂量反应线和样品的效价估计值。两种U-HuEPO制剂，即第2国际参考制剂和CAT-1，以及R-HuEPO被不同地用作标准品。测试的样品包括临床相关的人血清以及U-HuEPO和R-HuEPO的部分纯化制剂。在每个放射免疫分析以及测试的每个分析变体中，样品给出的剂量反应线的斜率与标准品的斜率无显著差异。对于两种变体放射免疫分析中的每一种，任何样品的效价估计值都是一致的，并且在进行检查时，放射免疫分析效价估计值与体内生物测定结果一致。在两个实验室独立获得的这些结果表明，使用R-HuEPO衍生试剂可以开发出对临床样品中EPO检测具有适当特异性和灵敏度的放射免疫分析。
