Lee C Y, Leung W Y, Tung J K
Biotechnol Appl Biochem. 1987 Feb;9(1):31-8.
Monoclonal antibodies to human liver ferritin were generated by an improved hybridoma technique using a semisolid medium containing methylcellulose for initial cloning after the cell fusion. Out of more than 1000 hybrid clones, only 1 was shown to secrete high-affinity monoclonal antibodies to human liver ferritin. The immunoglobulin subclass of this antibody was determined to be IgG2. The association constant between liver ferritin and this antibody was determined to be greater than 1 X 10(10) M-1. Due to the oligomeric nature of ferritin, this antibody can be simultaneously utilized as the first and second antibody in solid-phase sandwich immunoradiometric and enzyme immunoassays. This immunoassay procedure can be performed within 30-45 min and has a sensitivity of about 1 ng/ml. Under identical assay conditions, ferritin isolated from human spleen and human heart gave 50 and 30% cross-reactivity, respectively.
采用改良的杂交瘤技术，使用含有甲基纤维素的半固体培养基在细胞融合后进行初始克隆，从而制备出了抗人肝铁蛋白的单克隆抗体。在1000多个杂交克隆中，只有1个被证明能分泌抗人肝铁蛋白的高亲和力单克隆抗体。该抗体的免疫球蛋白亚类被确定为IgG2。肝铁蛋白与该抗体之间的结合常数被确定大于1×10(10) M-1。由于铁蛋白的寡聚性质，该抗体可同时用作固相夹心免疫放射测定法和酶免疫测定法中的第一抗体和第二抗体。该免疫测定程序可在30 - 45分钟内完成，灵敏度约为1 ng/ml。在相同的测定条件下，从人脾脏和人心脏分离的铁蛋白的交叉反应率分别为50%和30%。
