Lambert I, Kiełczyńska-Prekier K, Borysewicz G, Janicki A
Probl Med Wieku Rozwoj. 1975;5:57-65.
All the methods of quantitative estimation of transferrin concentration applied until recently have consisted in calculating the amount of this protein through the amount of iron bound in the blood serum. For this purpose the total iron binding capacity of serum (TIBC) has to be determined and then transferrin concentration is calculated. The calculation is based on the fact found by Ehrenberg and Laurell (1955) that two atoms of trivalent iron are selectively bound with one molecule of transferrin. The disadvantage of this method consists mainly in the necessity to carry out the measurement twice in every serum sample and in the large amount of material necessary for the analyses: this is a great inconvenience for pediatric clinics. The single-radial diffusion method in agar-gel was evaluated to be very advantageous for quantitative estimation of transferrin because of its simple technique and reaction specifity. The quantitative estimation is possible by the pilot calibration curve that represents the relationship of the diameter of a given circle and transferrin concentration. The quantity of material necessary for analysis amounts to not more than 5 mul of serum. Specific antitransferrin serum has to be imported and this seems to be the only reason why the said technique has not been used in Poland so far. The sera and vaccine laboratories in Warsaw have recently started to produce this antiserum and pure human transferrin. This has stimulated us to work out the single-radial diffusion method in agar based on Polish antiserum and Polish standard. The quality of the Polish specific rabbit antitransferrin serum and Polish pure human transferrin was tested by the immunoelectrophoretic method. The above findings provide the evidence that the Polish reagents are as good as those of the Behringwerke. Single-radial diffusion in agar gel should be carried out under the following conditions. 1) reaction temperature: 20-22degreesC 2) reaction time 24 hours 3) optimum concentration of antitransferrin serum in agar is 1:5.2. The error of the immunochemical method was determined experimentally and was found to be 9.05%. The results obtained with Polish and Behringwerke reagents were compared and were very similar.
直到最近,所有用于定量估算转铁蛋白浓度的方法都是通过血清中结合铁的量来计算这种蛋白质的含量。为此,必须先测定血清的总铁结合能力(TIBC),然后再计算转铁蛋白浓度。该计算基于埃伦贝格和劳雷尔(1955年)发现的事实,即两个三价铁原子与一个转铁蛋白分子选择性结合。这种方法的缺点主要在于每个血清样本都需要进行两次测量,且分析所需的材料量很大:这对儿科诊所来说非常不便。由于其技术简单和反应特异性,琼脂凝胶单向扩散法被评估为对转铁蛋白进行定量估算非常有利。通过代表给定圆圈直径与转铁蛋白浓度关系的初步校准曲线可以进行定量估算。分析所需的材料量不超过5微升血清。必须进口特异性抗转铁蛋白血清,这似乎是到目前为止波兰尚未使用该技术的唯一原因。华沙的血清和疫苗实验室最近开始生产这种抗血清和纯人转铁蛋白。这促使我们基于波兰抗血清和波兰标准开发琼脂单向扩散法。通过免疫电泳法检测了波兰特异性兔抗转铁蛋白血清和波兰纯人转铁蛋白的质量。上述结果证明波兰试剂与贝林公司的试剂一样好。琼脂凝胶单向扩散应在以下条件下进行。1)反应温度:20 - 22摄氏度;2)反应时间:24小时;3)琼脂中抗转铁蛋白血清的最佳浓度为1:5.2。通过实验确定了免疫化学方法的误差为9.05%。比较了使用波兰试剂和贝林公司试剂获得的结果,发现二者非常相似。