通过细胞内内含肽介导的蛋白质反式剪接产生骆驼科VHH双特异性构建体。
Generation of camelid VHH bispecific constructs via in-cell intein-mediated protein trans-splicing.
作者信息
Shibuya Yuki, Haga Natsuki, Asano Ryutaro, Nakazawa Hikaru, Hattori Takamitsu, Takeda Daisuke, Sugiyama Aruto, Kurotani Reiko, Kumagai Izumi, Umetsu Mitsuo, Makabe Koki
机构信息
Graduate School of Science and Engineering, Yamagata University, 4-3-16 Jyonan, Yonezawa, Yamagata 992-8510, Japan.
Department of Biotechnology and Life Science, Graduate School of Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 183-8538, Japan.
出版信息
Protein Eng Des Sel. 2017 Jan;30(1):15-21. doi: 10.1093/protein/gzw057. Epub 2016 Nov 23.
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.
生产仅重链抗体（VHH）重链双特异性可变结构域的各种组合以评估其治疗潜力通常需要几个基因工程步骤。在此，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构建体的替代方法；该方法可能会减少所需的基因操作步骤数量。作为概念验证，我们构建了一种包含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），并评估和确认了其双特异性。我们还使用PTS反应通过荧光蛋白标记测试了抗体标记。与传统的基因构建方法相比，通过PTS构建bsAb是生成多种bsAb组合的一种有前景的替代方法。
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