ANTIBODIES TO HUMAN TYPE IV COLLAGENASE & ITS INHIBITOR

基底膜的降解对白细胞的增殖具有重要作用 迁移、组织重塑、伤口愈合、血管生成和肿瘤 入侵IV型胶原是基底膜的主要成分， 由IV型胶原酶（CIVase）降解，CIVase通常存在于 潜伏形式潜伏的CIV酶可以在体内被激活为活性的CIV酶。 其活性可由内源性组织抑制剂调节， Ⅱ型金属蛋白酶（TIMP 2）。到目前为止，还没有具体的 商业上基于生物、酶或抗体的诊断试剂盒 可用于测定CIVase或TIMP 2，具有可接受的灵敏度。我们 最近已经产生了针对CIVase的单克隆抗体（mAb）。 这些单克隆抗体已通过免疫沉淀，Western印迹， CIVase的免疫细胞化学标记。我们将使用类似的 产生抗TIMP 2 mAb的方法，并使用这些mAb开发 定量酶联免疫吸附测定（ELISA）， 免疫组化和流式细胞术检测CIVase， TIMP 2在人体体液和组织中的水平。在第二阶段，我们将 使用这些mAb评价CIVase和 疾病的预后水平，包括 基底膜我们还将分离编码这些抗- CIVase和抗TIMP 2 mAb显示中和活性以产生 单链抗原结合蛋白或人源化嵌合抗体。 这些分子将被测试用于伤口的潜在治疗用途。 愈合、血管生成、类风湿性关节炎的治疗和预防 癌症转移的可能性

The degradation of basement membrane is important for leukocyte migration, tissue remodeling, wound healing, angiogenesis and tumor invasion. Type IV collagen, the major component of basement membrane, is degraded by type IV collagenase (CIVase) which usually exists in a latent form. The latent CIVase can be activated in vivo to an active form whose activity can be regulated by endogenous tissue inhibitor of metalloproteinase type II (TIMP2). As yet there is no specific biological, enzymatic or antibody based diagnostic kit commercially available with acceptable sensitivity for assaying CIVase or TIMP2. We have recently generated monoclonal antibodies (mAbs) against CIVase. These mAbs have been characterized by immunoprecipitation, Western blot, and immunocytochemical labeling of CIVase. We will use a similar approach to generate anti-TIMP2 mAbs and use these mAbs to develop quantitative enzyme-linked immunosorbent assay (ELISA), immunohistochemical and flow cytometry test for examining CIVase and TIMP2 level in the human body fluids and tissues. In phase II, we will use these mAbs to evaluate clinical correlations between CIVase and prognosis levels of patients with diseases involving the breakdown of basement membrane. We will also isolate cDNA coding for these anti- CIVase and anti-TIMP2 mAbs that show neutralization activity to generate single chain antigen-binding proteins or humanized chimeric antibodies. These molecules will be tested for potential therapeutic uses in wound healing, angiogenesis, treatment of rheumatoid arthritis and prevention of cancer metastasis.