开发新的基因型-表型关联抗体筛选系统。

IF 6.4 1区 生物学 Q1 BIOLOGY
eLife Pub Date : 2024-11-19 DOI:10.7554/eLife.95346
Takashi Watanabe, Hikaru Hata, Yoshiki Mochizuki, Fumie Yokoyama, Tomoko Hasegawa, Naveen Kumar, Tomohiro Kurosaki, Osamu Ohara, Hidehiro Fukuyama
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引用次数: 0

摘要

抗体是治疗和诊断各种疾病的有力工具。除了传统的基于杂交瘤的筛选外,基于重组抗体的筛选已成为一种常见的选择;然而,其应用受到两个因素的阻碍:(1) 筛选始于 Ig 基因克隆和重组抗体生产之后;(2) 抗体由成对的重链和轻链组成,通常由两个独立的表达载体表达。在这里,我们介绍了一种快速筛选重组单克隆抗体的方法,即建立基于黄金门的双表达载体,并在体内表达膜结合抗体。利用该系统,我们展示了在 7 天内从免疫小鼠体内快速分离出具有高亲和力的流感交叉反应抗体的方法。该系统特别适用于分离治疗或诊断抗体,例如在可预见的大流行期间。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Development of a new genotype-phenotype linked antibody screening system.

Antibodies are powerful tools for the therapy and diagnosis of various diseases. In addition to conventional hybridoma-based screening, recombinant antibody-based screening has become a common choice; however, its application is hampered by two factors: (1) screening starts after Ig gene cloning and recombinant antibody production only, and (2) the antibody is composed of paired chains, heavy and light, commonly expressed by two independent expression vectors. Here, we introduce a method for the rapid screening of recombinant monoclonal antibodies by establishing a Golden Gate-based dual-expression vector and in-vivo expression of membrane-bound antibodies. Using this system, we demonstrate the rapid isolation of influenza cross-reactive antibodies with high affinity from immunized mice within 7 days. This system is particularly useful for isolating therapeutic or diagnostic antibodies, for example during foreseen pandemics.

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来源期刊
eLife
eLife BIOLOGY-
CiteScore
12.90
自引率
3.90%
发文量
3122
审稿时长
17 weeks
期刊介绍: eLife is a distinguished, not-for-profit, peer-reviewed open access scientific journal that specializes in the fields of biomedical and life sciences. eLife is known for its selective publication process, which includes a variety of article types such as: Research Articles: Detailed reports of original research findings. Short Reports: Concise presentations of significant findings that do not warrant a full-length research article. Tools and Resources: Descriptions of new tools, technologies, or resources that facilitate scientific research. Research Advances: Brief reports on significant scientific advancements that have immediate implications for the field. Scientific Correspondence: Short communications that comment on or provide additional information related to published articles. Review Articles: Comprehensive overviews of a specific topic or field within the life sciences.
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