在人肿瘤异种移植模型中对靶向结合进行直接体内测量。
Direct in vivo measurement of targeted binding in a human tumor xenograft.
作者信息
Berk D A, Yuan F, Leunig M, Jain R K
机构信息
Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston 02114, USA.
出版信息
Proc Natl Acad Sci U S A. 1997 Mar 4;94(5):1785-90. doi: 10.1073/pnas.94.5.1785.
Binding is crucial to the function of most biologically active molecules, but difficult to quantify directly in living tissue. To this end, fluorescence recovery after photobleaching was used to detect the immobilization of fluorescently labeled ligand caused by binding to receptors in vivo. Measurements of mAb affinity to target antigen within human tumor xenografts revealed a saturable binding isotherm, from which an in vivo carcinoembryonic antigen density of 0.56 nmol/g (5.0 x 10(5)/cell) and an association constant of Ka < or = 4 x 10(7) M(-1) were estimated. The present method can be adapted for in vivo studies of cell signaling, targeted drugs, gene therapy, and other processes involving receptor-ligand binding.
结合对于大多数生物活性分子的功能至关重要，但在活体组织中直接定量却很困难。为此，采用光漂白后荧光恢复技术来检测体内荧光标记配体与受体结合导致的固定化。对人肿瘤异种移植体内单克隆抗体与靶抗原亲和力的测量揭示了一个可饱和的结合等温线，据此估计体内癌胚抗原密度为0.56 nmol/g（5.0×10⁵/细胞），缔合常数Ka≤4×10⁷ M⁻¹。本方法可适用于细胞信号传导、靶向药物、基因治疗以及其他涉及受体 - 配体结合过程的体内研究。
Zotero 插件