纤维连接蛋白降解片段(MAD2)单抗的制备及肝细胞癌患者血浆含量检测

PREPARATION OF MONOCLONAL ANTIBODY AGAINST THE DEGRADATION FRAGMENT OF FIBRONECTIN (MAD2)AND DETECTION OF SERUM MAD2 IN THE PATIENTS WITH HEPATOCELLULAR CARCINOMA

应用杂交瘤技术制备了纤维连接蛋白 (FN)降解片段MAD2的单克隆抗体 (McAb) ;建立了MAD2检测的双抗体夹心ELISA法 ;对 2 2 7例肝细胞癌 (HCC)、76例肝转移癌 (HMC)、98例消化道癌 (ACC)、15 6例慢性肝病 (CLD)患者和 48例健康人体血浆MAD2含量进行测定。结果显示 ,制备的MAD2McAb属IgG1,与FN无交叉反应 ;HCC组血浆MAD2含量均值与CLD组、HMC组、ACC组和正常对照组比较差异有显著性意义 (分别P <0 0 1) ,以HCC患者的最低MAD2值作临界值时 ,仅 13 5 %CLD、6 6 %HMC和 4 1%ACC超过此值 ,而健康人体血浆MAD2含量均低于临界值 ;6 5例血清AFP正常的HCC患者其血浆平均MAD2值仍明显高于非HCC肿瘤、CLD患者和正常对照。结合以前的结果 ,进一步表明 ,血浆MAD2检测可作为HCC诊断的新标志物 ,并可与AFP相互补充 ;MAD2 McAb的制备使MAD2的检测变得易行。

In order to develop ELISA method for measuring the degradation fragment of fibronectin (MAD2),hybridoma technique was used to obtain the monoclonal antibody (McAb)IgG 1 against MAD2 without cross reaction with fibronectin.The sera from 277 patients with hepatocellular carcinoma (HCC),76 with metastatic hepatic carcinoma (HMC),98 with alimentary canal carcinoma (ACC) and 156 with chronic liver disease(CLD) and 48 healthy subjects were assayed with ELISA method using this antibody.The examination showed that the mean value of MAD2 from the patients with HCC showed obviously significant difference compared with those of CLD,HMC,ACC and normal control groups ( p <0 01,respectively).If the lowest MAD2 value of HCC group was used as the cutoff value(CV).It was found that only 13 5% of CLD,6 6% of HMC,4 1% of ACC and 0 0% of normal controls were over the cv.Meanwhile,65 patients with HCC whose serum AFP levels were normal had also higher mean level of serum MAD2 than those of non HCC groups. The above results strongly suggested that serum MAD2 may be a new sensitive indicator for the diagnosis of patients with HCC.The preparation of MAD2 McAb makes the direct examination of MAD2 possible.