Improved stability of multivalent antibodies containing the human collagen XV trimerization domain
Improved stability of multivalent antibodies containing the human collagen XV trimerization domain
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DOI：
10.4161/mabs.4.2.19140
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发表时间：
2012-03-01
期刊：
影响因子：
5.3
通讯作者：
Alvarez-Vallina, Luis
中科院分区：
文献类型：
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作者：
Cuesta, Angel M.;Sanchez-Martin, David;Alvarez-Vallina, Luis
我们最近描述了一种工程化同源三聚抗体的体外和体内特性，该抗体是通过将胶原蛋白 XVIII NC1 结构域的 N 端三聚化区域融合到 scFv 片段的 C 端而制成的 [三聚体 (scFv-NC1)(3)； 110 kDa]。在这里，我们展示了胶原蛋白 XV NC1 结构域的 N 端三聚化区域在三价抗体工程中的实用性。我们构建了几种含有人XV型三聚结构域的基于scFv的三聚体，并证明所有纯化的三聚体在溶液中都是三聚体，并表现出优异的抗原结合能力。重要的是，XV型三聚体表现出比XVIII型三聚体显着更高的热稳定性和血清稳定性以及对蛋白酶消化的抗性。总之，XV 型胶原三聚结构域的小尺寸、高表达水平、溶解性和稳定性使其成为工程同源三聚抗体用于癌症检测和治疗的理想选择。
We recently described the in vitro and in vivo properties of an engineered homotrimeric antibody made by fusing the N-terminal trimerization region of collagen XVIII NC1 domain to the C-terminus of a scFv fragment [trimerbody(scFv-NC1)(3); 110 kDa]. Here, we demonstrated the utility of the N-terminal trimerization region of collagen XV NC1 domain in the engineering of trivalent antibodies. We constructed several scFv-based trimerbodies containing the human type XV trimerization domain and demonstrated that all the purified trimerbodies were trimeric in solution and exhibited excellent antigen binding capacity. Importantly, type XV trimerbodies demonstrated substantially greater thermal and serum stability and resistance to protease digestion than type XVIII trimerbodies. In summary, the small size, high expression level, solubility and stability of the trimerization domain of type XV collagen make it the ideal choice for engineering homotrimeric antibodies for cancer detection and therapy.