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一种通过琼脂糖天然凝胶电泳和接触印迹相结合来表征构象特异性抗体的新方法。

A New Method to Characterize Conformation-Specific Antibody by a Combination of Agarose Native Gel Electrophoresis and Contact Blotting.

作者信息

Akuta Teruo, Maruyama Toshiaki, Sakuma Chiaki, Nakagawa Masataka, Tomioka Yui, Entzminger Kevin, Fleming Jonathan K, Sato Ryo, Shibata Takashi, Kurosawa Yasunori, Okumura C J, Arakawa Tsutomu

机构信息

Research and Development Division, Kyokuto Pharmaceutical Industrial Co., Ltd., 3333-26, Aza-Asayama, Kamitezuna, Takahagi-shi 318-0004, Ibaraki, Japan.

Abwiz Bio Inc., 9823 Pacific Heights Blvd., Suite J, San Diego, CA 92121, USA.

出版信息

Antibodies (Basel). 2022 May 12;11(2):36. doi: 10.3390/antib11020036.

Abstract

In this study, we review the agarose native gel electrophoresis that separates proteins and macromolecular complexes in their native state and transfer of the separated proteins from the agarose gel to membranes by contact blotting which retains the native state of these structures. Green fluorescent protein showed functional state both on agarose gel and blotted membrane. Based on the combined procedures, we discovered conformation-specific monoclonal antibodies against PLXDC2 and SARS-CoV-2 spike protein.

摘要

在本研究中,我们回顾了琼脂糖原生凝胶电泳,它能分离处于天然状态的蛋白质和大分子复合物,以及通过接触印迹法将分离出的蛋白质从琼脂糖凝胶转移到膜上,这种方法能保留这些结构的天然状态。绿色荧光蛋白在琼脂糖凝胶和印迹膜上均显示出功能状态。基于这些组合程序,我们发现了针对PLXDC2和新冠病毒刺突蛋白的构象特异性单克隆抗体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4f69/9149980/d5bb9ae43244/antibodies-11-00036-g001.jpg

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