A multiplatform strategy for the discovery of conventional monoclonal antibodies that inhibit the voltage-gated potassium channel Kv1.3.
A multiplatform strategy for the discovery of conventional monoclonal antibodies that inhibit the voltage-gated potassium channel Kv1.3.
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DOI：
10.1080/19420862.2018.1445451
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发表时间：
2018
期刊：
影响因子：
5.3
通讯作者：
Colussi P
中科院分区：
文献类型：
--
作者：
Bednenko J;Harriman R;Mariën L;Nguyen HM;Agrawal A;Papoyan A;Bisharyan Y;Cardarelli J;Cassidy-Hanley D;Clark T;Pedersen D;Abdiche Y;Harriman W;van der Woning B;de Haard H;Collarini E;Wulff H;Colussi P
鉴定阻断人电压门控离子通道（VGIC）的单克隆抗体是一项具有挑战性的奋进，由于难以以支持药物发现计划的量生产重组离子通道蛋白而加剧。我们已经开发了一种通用策略来解决这一挑战，通过将四膜虫中重组VGIC的高水平表达与遗传多样性物种的免疫接种和独特的筛选工具相结合，从而深入挖掘可能结合蛋白质功能重要区域的抗体。使用这种方法，我们靶向人Kv1.3，一种电压门控钾通道，被广泛认为是治疗各种T细胞介导的自身免疫性疾病的治疗靶点。用重组Kv1. 3从免疫鸡和美洲驼中制备并回收了69株全长抗Kv1.3单抗，其中10株能抑制Kv1. 3电流。选择的抗体显示出对Kv1.3的效力（IC50<10 nM）和特异性超过相关的Kv1家族成员hERG和hNav1.5。
Identifying monoclonal antibodies that block human voltage-gated ion channels (VGICs) is a challenging endeavor exacerbated by difficulties in producing recombinant ion channel proteins in amounts that support drug discovery programs. We have developed a general strategy to address this challenge by combining high-level expression of recombinant VGICs in Tetrahymena thermophila with immunization of phylogenetically diverse species and unique screening tools that allow deep-mining for antibodies that could potentially bind functionally important regions of the protein. Using this approach, we targeted human Kv1.3, a voltage-gated potassium channel widely recognized as a therapeutic target for the treatment of a variety of T-cell mediated autoimmune diseases. Recombinant Kv1.3 was used to generate and recover 69 full-length anti-Kv1.3 mAbs from immunized chickens and llamas, of which 10 were able to inhibit Kv1.3 current. Select antibodies were shown to be potent (IC50<10 nM) and specific for Kv1.3 over related Kv1 family members, hERG and hNav1.5.
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影响因子：
10.8
作者：
Cheong A;Li J;Sukumar P;Kumar B;Zeng F;Riches K;Munsch C;Wood IC;Porter KE;Beech DJ
通讯作者：
Beech DJ
DOI：
10.1152/ajprenal.00374.2010
发表时间：
2010-12-01
影响因子：
4.2
作者：
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通讯作者：
Kumagai, Hiroo
影响因子：
4.8
作者：
de Haard, HJ;van Neer, N;Hoogenboom, HR
通讯作者：
Hoogenboom, HR
影响因子：
5.3
作者：
Blomberg, P;Randolph, C;Yao, MC
通讯作者：
Yao, MC
影响因子：
2.9
作者：
Helms, LMH;Felix, JP;Slaughter, RS
通讯作者：
Slaughter, RS