Brophy C M, Marks W H, Reilly J M, Tilson M D
Department of Surgery, Yale University School of Medicine, New Haven, CT 06510.
Biochem Biophys Res Commun. 1990 Mar 30;167(3):898-903. doi: 10.1016/0006-291x(90)90608-p.
Tissue inhibitor of metalloproteinases (TIMP) is the major inhibitor of collagenase, gelatinase, proteoglycanase, stromelysin, and metalloelastases. An imbalance between proteases and inhibitors has been implicated in numerous disease processes including tumor invasion, rheumatoid arthritis, emphysema, and aortic aneurysm disease. The purpose of this investigation was to develop a polyclonal antibody to recombinant TIMP and establish an immunoassay to measure immunoreactive protein in normal and diseased tissues. A polyclonal antibody was produced in rabbit against recombinant human TIMP which was characterized and used to establish a radioimmunoassay. The assay was used to measure immunoreactive protein in fibroblast conditioned medium, human serum, and aortic extracts. There was more immunoreactive TIMP in matrix associated urea extracts than soluble salt extracts from human aorta, suggesting that TIMP is matrix associated. The sensitivity of the assay enables the specific measurement of this inhibitor in serum, fibroblast culture medium, and tissue extracts.
金属蛋白酶组织抑制剂（TIMP）是胶原酶、明胶酶、蛋白聚糖酶、基质溶解素和金属弹性蛋白酶的主要抑制剂。蛋白酶和抑制剂之间的失衡与许多疾病过程有关，包括肿瘤侵袭、类风湿性关节炎、肺气肿和主动脉瘤疾病。本研究的目的是开发一种针对重组TIMP的多克隆抗体，并建立一种免疫测定法来测量正常和患病组织中的免疫反应性蛋白。用兔制备了针对重组人TIMP的多克隆抗体，对其进行了表征并用于建立放射免疫测定法。该测定法用于测量成纤维细胞条件培养基、人血清和主动脉提取物中的免疫反应性蛋白。与来自人主动脉的可溶性盐提取物相比，基质相关尿素提取物中存在更多的免疫反应性TIMP，这表明TIMP与基质相关。该测定法的灵敏度能够特异性测量血清、成纤维细胞培养基和组织提取物中的这种抑制剂。
