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

IF 3 Q3 IMMUNOLOGY

Antibodies Pub Date : 2022-05-12 DOI:10.3390/antib11020036

T. Akuta, T. Maruyama, Chiaki Sakuma, Masataka Nakagawa, Yui Tomioka, Kevin C Entzminger, J. Fleming, Ryo Sato, Takashi Shibata, Y. Kurosawa, C. Okumura, T. Arakawa

引用次数: 4

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.

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琼脂糖天然凝胶电泳与接触印迹相结合的构象特异性抗体鉴定新方法

在这项研究中，我们回顾了琼脂糖天然凝胶电泳，该电泳分离天然状态的蛋白质和大分子复合物，并通过接触印迹将分离的蛋白质从琼脂糖凝胶转移到膜上，从而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表现出功能状态。基于联合程序，我们发现了针对PLXDC2和严重急性呼吸系统综合征冠状病毒2型刺突蛋白的构象特异性单克隆抗体。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Antibodies IMMUNOLOGY-

CiteScore

7.10

自引率

6.40%

发文量

审稿时长

11 weeks

期刊介绍： Antibodies (ISSN 2073-4468), an international, peer-reviewed open access journal which provides an advanced forum for studies related to antibodies and antigens. It publishes reviews, research articles, communications and short notes. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. Full experimental and/or methodical details must be provided. Electronic files or software regarding the full details of the calculation and experimental procedure - if unable to be published in a normal way - can be deposited as supplementary material. This journal covers all topics related to antibodies and antigens, topics of interest include (but are not limited to): antibody-producing cells (including B cells), antibody structure and function, antibody-antigen interactions, Fc receptors, antibody manufacturing antibody engineering, antibody therapy, immunoassays, antibody diagnosis, tissue antigens, exogenous antigens, endogenous antigens, autoantigens, monoclonal antibodies, natural antibodies, humoral immune responses, immunoregulatory molecules.