Su J L, Becherer J D, Edwards C, Bukhart W, McGeehan G M, Champion B R
Department of Cell Physiology, Glaxo Inc. Research Institute, Research Triangle Park, North Carolina 27709, USA.
Hybridoma. 1995 Aug;14(4):383-90. doi: 10.1089/hyb.1995.14.383.
Mouse monoclonal antibodies against recombinant human fibroblast procollagenase and prostromelysin have been generated and characterized. The epitope-containing domains for the antibodies have been assigned based on their immunoreactivities against recombinant proenzymes, mature enzymes, truncated collagenases, proteolytic fragments of stromelysin, and chimeric molecules constructed from different domains of the two enzymes. These antibodies can be divided into four groups: (1) antibodies that recognize the truncated 19-kDa NH2-terminal collagenase, (2) antibodies that recognize the C-terminal domain of collagenase and stromelysin, (3) antibodies that recognize a 31-kDa NH2-terminal collagenase fragment, and (4) antibodies that recognize the 19-kDa NH2-fragment of stromelysin. The prostromelysin-specific antibody 11N13 is of particular interest; it neutralizes stromelysin activity in a stromelysin peptide substrate assay, with an IC50 value of 75 nM. MAb 11N13 may be useful for in vivo and in vitro studies to validate the roles of stromelysin in tumor cell invasion, metastasis, and connective tissue disorders.
已制备并鉴定了针对重组人成纤维细胞原胶原酶和原基质溶解素的小鼠单克隆抗体。基于这些抗体对重组酶原、成熟酶、截短的胶原酶、基质溶解素的蛋白水解片段以及由这两种酶的不同结构域构建的嵌合分子的免疫反应性，确定了抗体的表位结构域。这些抗体可分为四组：(1) 识别截短的19 kDa NH2末端胶原酶的抗体；(2) 识别胶原酶和基质溶解素C末端结构域的抗体；(3) 识别31 kDa NH2末端胶原酶片段的抗体；(4) 识别基质溶解素19 kDa NH2片段的抗体。原基质溶解素特异性抗体11N13特别受关注；在基质溶解素肽底物测定中，它可中和基质溶解素活性，IC50值为75 nM。单克隆抗体11N13可能有助于体内和体外研究，以验证基质溶解素在肿瘤细胞侵袭、转移和结缔组织疾病中的作用。
