Zucker S, Lysik R M, Gurfinkel M, Zarrabi M H, Stetler-Stevenson W, Liotta L A, Birkedal-Hansen H, Mann W
Department of Research, Department of Veterans Affairs Medical Center, Northport, NY 11768.
J Immunol Methods. 1992 Apr 8;148(1-2):189-98. doi: 10.1016/0022-1759(92)90172-p.
We have developed a sensitive and specific sandwich type enzyme-linked immunosorbent assay (ELISA) to detect type IV collagenase (MMP-2) in human plasma which employs the combination of affinity purified rabbit polyclonal antibodies and mouse monoclonal antibodies to human MMP-2. The MMP-2 ELISA detects latent and activated MMP-2, MMP-2 complexed with TIMP and MMP-2 complexed with alpha 2 macroglobulin (65% efficiency). To determine whether physiologic conditions associated with increased connective tissue turnover are accompanied by increased MMP-2 levels in plasma, we compared enzyme levels in pregnant and nonpregnant women. Plasma MMP-2 (mean +/- standard deviation) was significantly increased (p less than 0.05) in the second half of pregnancy (650 +/- 312) as compared to early pregnancy (356 +/- 139) or the nonpregnant state (387 +/- 193). As a result of the linkage between type IV collagenase production by cancer cells and the metastatic phenotype, the assay of MMP-2 in plasma is of potential clinical value in cancer.
我们开发了一种灵敏且特异的夹心型酶联免疫吸附测定法(ELISA),用于检测人血浆中的IV型胶原酶(MMP - 2),该方法采用了亲和纯化的兔多克隆抗体和抗人MMP - 2小鼠单克隆抗体的组合。MMP - 2 ELISA可检测潜伏型和活化型MMP - 2、与组织金属蛋白酶抑制剂(TIMP)复合的MMP - 2以及与α2巨球蛋白复合的MMP - 2(效率为65%)。为了确定与结缔组织更新增加相关的生理状况是否伴随着血浆中MMP - 2水平的升高,我们比较了孕妇和非孕妇的酶水平。与妊娠早期(356±139)或非妊娠状态(387±193)相比,妊娠后半期血浆MMP - 2(平均值±标准差)显著升高(p<0.05)(650±312)。由于癌细胞产生IV型胶原酶与转移表型之间存在联系,血浆中MMP - 2的检测在癌症方面具有潜在的临床价值。