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由于狄戈辛的治疗浓度和中毒浓度很接近,需要快速检出血药浓度以指导用药。常用的放射免疫(RIA)方法分离和洗涤程序繁琐,临床应用不方便。作者根据待分离分子表面性质的差异,利用不溶混性的水-聚合物二相系统直接分离生物分子,从而大大简化了分离步骤。此法称为分溶亲和配基试验(Partition AffinityLigand Assay,简称PALA)。测定方法:将50μl血清与50μl~(125)I标记的狄戈辛混合,加入100μl抗狄戈辛抗体(游离或与葡聚糖结合),3～5分钟后加入预先混合好的聚合物系统[1ml 30%(W/W)聚乙二醇(PEG4000)和3ml30%(W/W)MgSO_4·7H_2O]并振摇,几分钟后二相分开,吸取上层PEG疏水相500μl进行计数,从标准曲线即可求得血清狄戈辛含量。整个试验不到10分种。
Due to the therapeutic concentration of Digoxin and the concentration of poisoning is very close, the need for rapid detection of blood concentration to guide the medication. Commonly used radioimmunoassay (RIA) method of separation and washing procedures cumbersome, inconvenient clinical application. Based on the differences in the surface properties of the molecules to be separated, the authors directly separate the biomolecules using an immiscible water-polymer two-phase system, thereby greatly simplifying the separation step. This method is called the Partition Affinity Ligand Assay (PALA for short). Assay Method: 50 μl of serum was mixed with 50 μl of 125 I-labeled Digoxin, 100 μl of anti-Digoxigenin antibody (either free or bound to dextran) was added, and added to the premixed polymer system after 3 to 5 minutes [1 ml of 30% w / w polyethylene glycol (PEG 4000) and 3 ml of 30% w / w MgSO 4 .7H 2 O] and shaken. Two minutes later, the two phases were separated and 500 μl of the upper PEG hydrophobic phase was aspirated for counting. Curves can be obtained from serum Digoxigenin content. The whole experiment less than 10 minutes.