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.

鉴定能阻断人电压门控离子通道（VGIC）的单克隆抗体是一项艰巨的挑战，因为难以生产支持药物发现程序的重组离子通道蛋白。我们已经开发了一种通用策略，通过结合在嗜热四膜膜虫中重组VGIC的高水平表达来应对这一挑战通过对系统发育多样的物种进行免疫接种，以及独特的筛选工具，可深入挖掘可能与蛋白质功能重要区域结合的抗体。使用这种方法，我们靶向人Kv1.3，这是一种电压门控钾离子通道，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标。重组Kv1.3用于从免疫鸡和美洲驼中生成和回收69种全长抗Kv1.3单克隆抗体，其中10种能够抑制Kv1.3电流。所选抗体显示出有力的（IC ₅₀ <10 nM），并且对相关的Kv1家族成员hERG和hNav1.5具有Kv1.3特异性。