Mahoney D P, Barden A, Beilin L J, Vandongen R
Prostaglandins Leukot Med. 1983 Sep;12(1):11-20. doi: 10.1016/0262-1746(83)90063-x.
A comparison of radioimmunoassay (R.I.A.) for 6-keto PGF 1 alpha, PGE2 and TXB2 using 125I-histamine ligands and tritiated tracers showed a 7 to 10 fold increase in sensitivity for the 125I derivative, with a resultant reduction in antisera requirements by 80%. Study of the effect of purification of biological samples by organic extraction and thin layer chromatography showed that for human and rat urine, and urine from perfused rat kidneys, direct R.I.A. of unextracted samples gave substantially higher estimates of PGE2, 6-keto PGF1 alpha and TXB2 than R.I.A. purified material. Indomethacin pre-treatment reduced the levels estimated in both extracted and unextracted samples; the results indicating that the higher estimates obtained by direct assay were due to a combination of PG/TXB2 metabolites and non-specific interference. In serum from incubated human blood, estimated 6-keto PGF1 alpha and PGE2 levels were significantly higher by direct R.I.A. than after extraction, whereas TXB2 levels were similar. Thus purification procedures are unnecessary when high serum TXB2 levels are being measured by R.I.A. with relatively specific antisera.
使用¹²⁵I - 组胺配体和氚标记示踪剂对6 - 酮 - PGF₁α、PGE₂和TXB₂进行放射免疫测定（R.I.A.）的比较表明，¹²⁵I衍生物的灵敏度提高了7至10倍，抗血清需求量相应减少了80%。对通过有机萃取和薄层色谱法纯化生物样品的效果研究表明，对于人尿、大鼠尿以及灌注大鼠肾脏的尿液，未萃取样品的直接放射免疫测定对PGE₂、6 - 酮 - PGF₁α和TXB₂的估计值显著高于经放射免疫测定纯化后的物质。吲哚美辛预处理降低了萃取和未萃取样品中的估计水平；结果表明，直接测定获得的较高估计值是由于PG/TXB₂代谢产物和非特异性干扰共同作用的结果。在人血孵育血清中，直接放射免疫测定法估计的6 - 酮 - PGF₁α和PGE₂水平显著高于萃取后水平，而TXB₂水平相似。因此，当使用相对特异的抗血清通过放射免疫测定法测量高血清TXB₂水平时，纯化程序是不必要的。
