Transport across the primate blood-brain barrier of a genetically engineered chimeric monoclonal antibody to the human insulin receptor
Transport across the primate blood-brain barrier of a genetically engineered chimeric monoclonal antibody to the human insulin receptor
复制标题
DOI：
10.1023/a:1007592720793
复制
发表时间：
2000-03-01
影响因子：
3.7
通讯作者：
Pardridge, WM
中科院分区：
文献类型：
--
作者：
Coloma, MJ;Lee, HJ;Pardridge, WM
目的.脑药物靶向可以通过将通常不穿过血脑屏障（BBB）的药物缀合至脑药物递送载体来实现。鼠抗人胰岛素受体（human insulin receptor，HIR）单抗83-14是一种潜在的脑靶向药物载体，如果将该单抗基因工程化形成嵌合抗体，其中大部分免疫原性鼠序列被人抗体序列取代，方法.本研究描述了嵌合HIRMAb的基因的产生、蛋白质的表达和表征、用111-铟和125-碘放射性标记嵌合HIRMAb以及在静脉内施用[（111）In]嵌合HIRMAb后2小时进行的活灵长类动物脑的定量放射自显影。基于Western印迹和免疫放射测定，嵌合HIRMAb与鼠HIRMAb对靶抗原具有相同的亲和力，所述免疫放射测定使用从分泌可溶性HIR的CHO细胞系产生的无血清条件培养基中纯化的部分纯化的HIR亲和力。[I-125]嵌合HIRMAb与分离的人脑毛细血管紧密结合，这种结合被鼠HIRMAb阻断。将[（111）ln]嵌合HIRMAb静脉内给予麻醉的恒河猴，2小时脑扫描显示活灵长类动物脑对嵌合抗体的强烈摄取。已经产生了基因工程嵌合HIRMAb，并且嵌合抗体与原始鼠抗体对人和灵长类BBB HIR具有相同的反应性。这种嵌合HIRMAb可用于人类，用于通过BBB靶向通常不穿过BBB的神经诊断或神经治疗药物。
Purpose. Brain drug targeting may be achieved by conjugating drugs, that normally do not cross the blood-brain barrier (BBB), to brain drug delivery vectors. The murine 83-14 MAb to the human insulin receptor (HIR) is a potential brain drugs targeting vector that could be used in humans, if this MAb was genetically engineered to form a chimeric antibody, where most of the immunogenic murine sequences are replaced by human antibody sequence.,Methods. The present studies describe the production of the gene for the chimeric HIRMAb, expression and characterization of the protein, radiolabeling of the chimeric HIRMAb with 111-indium and 125-iodine, and quantitative autoradiography of living primate brain taken 2 hours after intravenous administration of the [(111)ln]chimeric HIRMAb.Results, The chimeric HIRMAb had identical affinity to the target antigen as the murine HIRMAb based on Western blotting and immunoradiometric assay using partially purified HIR affinity purified from serum free conditioned media produced by a CHO cell line secreting soluble HIR. The [I-125]chimeric HIRMAb was avidly bound to isolated human brain capillaries, and this binding was blocked by the murine HIRMAb. The [(111)ln]chimeric HIRMAb was administered intravenously to an anesthetized Rhesus monkey, and the 2 hour brain scan showed robust uptake of the chimeric antibody by the living primate brain.Conclusions. A genetically engineered chimeric HIRMAb has been produced, and the chimeric antibody has identical reactivity to the human and primate BBB HIR as the original murine antibody. This chimeric HIRMAb may be used in humans for drug targeting through the BBB of neurodiagnostic or neurotherapeutic drugs that normally do not cross the BBB.