通过细胞内蛋白介导的反式剪接产生骆驼类VHH双特异性构建体

Yukihiko Shibuya, N. Haga, R. Asano, H. Nakazawa, T. Hattori, D. Takeda, Aruto Sugiyama, R. Kurotani, I. Kumagai, M. Umetsu, K. Makabe
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引用次数: 8

摘要

生产重链或仅重链抗体(VHH)结构体的各种双特异性可变结构域组合以评估其治疗潜力通常需要几个基因工程步骤。在这里,我们提出了一种通过蛋白质反式剪接(PTS)反应在体内创建双特异性VHH构建物的替代方法;这种方法可以减少所需的基因操作步骤的数量。作为概念验证,我们构建了含有抗表皮生长因子受体VHH和抗绿色荧光蛋白VHH的双特异性抗体(bsAb),并对其双特异性进行了评估和确认。我们还使用PTS反应测试了荧光蛋白标记的抗体标记。与传统的基因构建方法相比,通过PTS构建bsAb是一种很有前途的生成多个bsAb组合的替代方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Generation of camelid VHH bispecific constructs via in-cell intein-mediated protein trans-splicing
Production of various combinations of bispecific variable domain of heavy chain of heavy chain-only antibody (VHH) constructs to evaluate their therapeutic potential usually requires several gene-engineering steps. Here, we present an alternative method of creating bispecific VHH constructs in vivo through protein trans-splicing (PTS) reaction; this method may reduce the number of gene manipulation steps required. As a proof-of-concept, we constructed a bispecific antibody (bsAb) containing an anti-epidermal growth factor receptor VHH and anti-green fluorescent protein VHH, and we evaluated and confirmed its bispecificity. We also tested antibody labeling by fluorescent protein tagging using the PTS reaction. Compared with the conventional gene construction method, bsAb construction via PTS is a promising alternative approach for generating multiple bsAb combinations.
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