摘要
本文采用CsCl等密度梯度离心法从裸鼠移植LS174t结肠癌组织匀浆液中提取肿瘤相关抗原。CsCl密度为1.45g/cm^3,离心为45 000r/min,15℃,39小时。在紫外280nm监测下可描出一个蛋白峰,用Abbe折射仪检测各收集管样品折光率换算出相应的密度值,从结果看,7、9管的浮力密度为1.44g/cm^3和1.49g/cm^3和文献报道基本一致。ELISA法进行免疫活性测定,提纯抗原,4株胰腺癌单抗反应亲和力最强在7、9管,2株结肠癌单抗反应亲和力最强在7、11管。以双抗夹心法胰腺癌单抗PS-9做检测抗体制备标准曲线,可看出肿瘤相关抗原在0.625μg/ml—20μg/ml范围,肿瘤相关抗原和OD值之间呈线性关系,相关系数0.9978。
A method of isolation tumor-associated antigens from colon carcinoma tissues with CsCl density-gradient centrifugation is reported. The density of CsCl is 1.45g/cm^3 and centrifugation performed at 15℃ for 39h at 45000 rpm. A protein peak could be detected by UV absorbance at 280nm. The collected samples are measured with Abbl refractometer and the density of samples are calculated. The densities in the 7th and 9th tubes are 1.448/cm^3 and 1.49g/cm^3 respectively. By ELISA assay, the highest immune reactivities of four anti-pancreatic carcinoma monoclonal antibodies lie in the 7th and 9th tubes. Two anti-colonic carcinoma monoclonal antibodies shown that the highest reactivities with the antigens were in the 7th and 11th tubes. On the standard curve, the detectable concentration of the tumor-associated antigen is in the range of 0.265μg/ml to 20μg/ml when measured with an anti-pancreatic cancer monoclonal antibody Ps-9 in double antibody sandwich ELISA. It shows a linear relativity between the concentration of tumor-associated antigen and OD value. The relative coefficient is 0.9978.
出处
《免疫学杂志》
CAS
CSCD
北大核心
1992年第2期129-132,共4页
Immunological Journal
关键词
肿瘤相关抗原
单克隆抗体
CsCl densitygradient centrifugation, Tumor associated antigen, Monoclonal antibody
