Imagawa M, Yoshitake S, Ishikawa E, Niitsu Y, Urushizaki I, Kanazawa R, Tachibana S, Nakazawa N, Ogawa H
Clin Chim Acta. 1982 Jun 3;121(3):277-89. doi: 10.1016/0009-8981(82)90237-6.
A highly sensitive sandwich enzyme immunoassay of human ferritin was developed. Polystyrene balls were coated with rabbit anti-human ferritin IgG by physical adsorption, and rabbit anti-human ferritin Fab' was purified by affinity chromatography and labelled with beta-D-galactosidase from Escherichia coli. Using the anti-ferritin-coated polystyrene balls and labelled anti-ferritin, the sensitivity obtained was 23 fg (0.05 amol) of ferritin per tube. The range of serum ferritin levels that could be determined using 0.1 microliter of serum was 0.23-4500 ng/ml, and even 2.3 pg/ml was measurable by using 10 microliter of serum. The coefficients of within-assay (n = 25) and between-assay (n = 10) variations were 5.9-8.8%. The regression equation and coefficient of correlation to a radioimmunoassay were Y(RIA) = 0.92 X(EIA) + 3.0 and 0.99 (n = 78), respectively. The corresponding sandwich radioimmunoassay was less sensitive, partly because the specific radioactivity of 125I-labelled anti-ferritin IgG used was not sufficiently high.
开发了一种高灵敏度的人铁蛋白夹心酶免疫测定法。通过物理吸附将兔抗人铁蛋白IgG包被在聚苯乙烯球上，通过亲和层析纯化兔抗人铁蛋白Fab'并用来自大肠杆菌的β-D-半乳糖苷酶进行标记。使用抗铁蛋白包被的聚苯乙烯球和标记的抗铁蛋白，每管获得的铁蛋白灵敏度为23 fg（0.05 amol）。使用0.1微升血清可测定的血清铁蛋白水平范围为0.23 - 4500 ng/ml，甚至使用10微升血清可检测到2.3 pg/ml。批内（n = 25）和批间（n = 10）变异系数分别为5.9 - 8.8%。与放射免疫测定法的回归方程和相关系数分别为Y(RIA) = 0.92 X(EIA) + 3.0和0.99（n = 78）。相应的夹心放射免疫测定法灵敏度较低，部分原因是所使用的125I标记抗铁蛋白IgG的比放射性不够高。
