An anti-transferrin receptor-avidin fusion protein exhibits both strong proapoptotic activity and the ability to deliver various molecules into cancer cells
An anti-transferrin receptor-avidin fusion protein exhibits both strong proapoptotic activity and the ability to deliver various molecules into cancer cells
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DOI：
10.1073/pnas.162362999
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发表时间：
2002-08-06
影响因子：
11.1
通讯作者：
Penichet, ML
中科院分区：
文献类型：
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作者：
Ng, PP;Dela Cruz, JS;Penichet, ML
我们开发了一种抗体融合蛋白（抗大鼠 TfR IgG3-Av），能够将不同的分子递送到癌细胞中。它由与大鼠转铁蛋白受体特异性的人 IgG3 的 C(H)3 区域基因融合的亲和素组成。它与生物素化分子（如葡萄糖氧化酶和 β-半乳糖苷酶）形成强烈的非共价相互作用，并通过受体介导的内吞作用将它们递送至大鼠骨髓瘤细胞系 Y3-Ag1.2.3 中。重要的是，β-半乳糖苷酶在内化后保留活性。此外，我们意外地发现抗大鼠TfR IgG3-Av，而非重组抗大鼠TfR IgG3或非特异性IgG3-Av，具有针对Y3-Ag1.2.3和大鼠T细胞淋巴瘤细胞系C58(NT)D.1.G.OVAR.1的促凋亡活性。在两种非造血谱系的大鼠细胞系（膀胱癌 BC47 和胶质肉瘤 9L）中未观察到这些活性。抗人 TfRIgG3-Avals 表现出针对人红白血病细胞系 K562 的促凋亡活性。研究表明，抗大鼠 TfR IgG3-Av 以二聚体形式存在，表明表面转铁蛋白受体的交联可能是细胞毒性活性的原因。这些发现表明，通过与抗生物素蛋白融合，可以将不表现出抑制活性的生长因子受体特异性抗体转化为对所选细胞具有显着内在细胞毒活性的药物。通过将生物素化治疗剂递送到癌细胞中可以增强抗肿瘤活性。这项技术的进一步发展可能会产生体内根除血液恶性肿瘤的有效疗法，以及在自体移植中离体清除癌细胞。
We have developed an antibody fusion protein (anti-rat TfR IgG3-Av) with the ability to deliver different molecules into cancer cells. It consists of avidin genetically fused to the C(H)3 region of a human IgG3 specific for the rat transferrin receptor. It forms strong, noncovalent interactions with biotinylated molecules such as glucose oxidase and beta-galactosidase, and delivers them into the rat myeloma cell line Y3-Ag1.2.3 through receptor-mediated endocytosis. Importantly, the beta-galactosidase retains activity after internalization. Furthermore, we have unexpectedly discovered that anti-rat TfR IgG3-Av, but not a recombinant anti-rat TfR IgG3 or a nonspecific IgG3-Av, possesses proapoptotic activities against Y3-Ag1.2.3 and the rat T cell lymphoma cell line C58 (NT) D.1.G.OVAR.1. These activities were not observed in two rat cell lines of nonhematopoietic lineage (bladder carcinoma BC47 and gliosarcoma 9L). Anti-human TfRIgG3-Avalsodemonstrated proapoptotic activity against the human erythroleukemia cell line K562. Studies showed that anti-rat TfR IgG3-Av exists as a dimer, suggesting that cross-linking of the surface transferrin receptor may be responsible for the cytotoxic activity. These findings demonstrate that it is possible to transform an antibody specific for a growth factor receptor that does not exhibit inhibitory activity into a drug with significant intrinsic cytotoxic activity against selected cells by fusing it with avidin. The antitumor activity may be enhanced by delivering biotinylated therapeutics into cancer cells. Further development of this technology may lead to effective therapeutics for in vivo eradication of hematological malignancies, and ex vivo purging of cancer cells in autologous transplantation.