Engineering and expression of a chimeric transferrin receptor monoclonal antibody for blood-brain barrier delivery in the mouse.
Engineering and expression of a chimeric transferrin receptor monoclonal antibody for blood-brain barrier delivery in the mouse.
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DOI：
10.1002/bit.22135
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发表时间：
2009-03-01
影响因子：
3.8
通讯作者：
Pardridge, William M.
中科院分区：
文献类型：
--
作者：
Boado, Ruben J.;Zhang, Yun;Wang, Yuntao;Pardridge, William M.
蛋白质治疗剂可以通过与血脑屏障（BBB）分子特洛伊木马的基因融合来递送穿过血脑屏障（BBB）。后者是针对BBB受体（例如胰岛素受体或转铁蛋白受体（TfR））的内源性肽或拟肽单克隆抗体（MAb）。融合蛋白已经用针对人胰岛素受体（HIR）的MAb工程化。然而，HIRMAb对啮齿动物胰岛素受体没有活性，并且不能用于穿过小鼠BBB的药物递送。针对小鼠TfR的大鼠8D3单克隆抗体在小鼠中作为药物递送系统是有活性的，并且本研究描述了大鼠8D3 TfRMAb的重链可变区（VH）和轻链可变区（VL）的克隆和测序。VH和VL分别与小鼠IgG 1重链和小鼠κ轻链的恒定区融合，以产生新的嵌合TfRMAb。用重链和轻链表达质粒双重转染COS细胞后，在COS细胞中表达嵌合TfRMAb，并通过蛋白G亲和层析纯化。使用放射受体测定和小鼠成纤维细胞，嵌合TfRMAb对鼠TfR的亲和力等于8D3 MAb。将嵌合TfRMAb放射性标记并注射到小鼠中用于嵌合TfRMAb清除的药代动力学研究。嵌合TfRMAb在体内以与大鼠8D3 MAb相当的水平被小鼠脑迅速摄取。总之，这些研究描述了对小鼠TfR具有高活性的嵌合TfRMAb的基因工程、表达和验证，其可用于小鼠中BBB药物递送的治疗性融合蛋白的未来工程。
Protein therapeutics may be delivered across the blood–brain barrier (BBB) by genetic fusion to a BBB molecular Trojan horse. The latter is an endogenous peptide or a peptidomimetic monoclonal antibody (MAb) against a BBB receptor, such as the insulin receptor or the transferrin receptor (TfR). Fusion proteins have been engineered with the MAb against the human insulin receptor (HIR). However, the HIRMAb is not active against the rodent insulin receptor, and cannot be used for drug delivery across the mouse BBB. The rat 8D3 MAb against the mouse TfR is active as a drug delivery system in the mouse, and the present studies describe the cloning and sequencing of the variable region of the heavy chain (VH) and light chain (VL) of the rat 8D3 TfRMAb. The VH and VL were fused to the constant region of mouse IgG1 heavy chain and mouse kappa light chain, respectively, to produce a new chimeric TfRMAb. The chimeric TfRMAb was expressed in COS cells following dual transfection with the heavy and light chain expression plasmids, and was purified by protein G affinity chromatography. The affinity of the chimeric TfRMAb for the murine TfR was equal to the 8D3 MAb using a radio-receptor assay and mouse fibroblasts. The chimeric TfRMAb was radio-labeled and injected into mice for a pharmacokinetics study of the clearance of the chimeric TfRMAb. The chimeric TfRMAb was rapidly taken up by mouse brain in vivo at a level comparable to the rat 8D3 MAb. In summary, these studies describe the genetic engineering, expression, and validation of a chimeric TfRMAb with high activity for the mouse TfR, which can be used in future engineering of therapeutic fusion proteins for BBB drug delivery in the mouse.
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影响因子：
11.4
作者：
DANGL, JL;WENSEL, TG;OI, VT
通讯作者：
OI, VT
影响因子：
3.8
作者：
Boado, Ruben J.;Zhang, Yufeng;Pardridge, William M.
通讯作者：
Pardridge, William M.
影响因子：
3.8
作者：
Boado, Ruben J.;Zhang, Yun;Pardridge, William M.
通讯作者：
Pardridge, William M.
影响因子：
2.3
作者：
Kissel, K;Hamm, S;Engelhardt, B
通讯作者：
Engelhardt, B
影响因子：
4.7
作者：
Boado, Ruben J.;Zhang, Yufeng;Pardridge, William M.
通讯作者：
Pardridge, William M.