肌醇磷脂激酶(PI4-K)单克隆抗体的制备及其对细胞生长的抑制

Devolopment of the Monoclonal Antibody Against Type Ⅱ Phosphatidylinositol 4-kinase from Bovine Cerebella Cortex and its Inhibitory Effect on Cell Growth

制备了抗肌醇磷脂激酶 ( PI4- K)单克隆抗体 ( A6D)并测定了抗原 -抗体反应基本特性及功能 .结果表明 ,单克隆抗体与固相及溶液中肌醇磷脂激酶的亲和常数分别为 7.5× 1 0 6和 6× 1 0 8( mol/L) -1.单抗 1 .9× 1 0 -7mol/L可以抑制从细胞提取液的 PI4- K酶活力 50 % .用 FITC标记单抗在蛋白微球引导下进入细胞内 ,主要富集在细胞质膜区 ,并对 He La细胞和小鼠小脑细胞生长有明显抑制作用 .

A monoclonal antibody (A6D) against phosphatidylinositol 4\|kinase(PI 4\|K)was developed and characterized.The results indicated that the binding affinity constants of the antibody to antigen (PI 4\|K)either coated or in solution were 7 5×10 6(mol/L) -1 and 6×10 8 (mol/L) -1 respectively.The FITC\|A6D conjugated with amphiphilic protein micro\|vesicular was located mainly in the plasma membrane of the HeLa cells observed by fluorescence microscopy.The PI 4 K activities could inhibit 50% by A6D of 2×10 -7 mol/L of cell lysates.The further study showed that the antibody could markedly inhibit the growth of both HeLa cells and mouse cerebella granule cells.