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新型基因型-表型关联抗体筛选系统的开发。

Development of a new genotype-phenotype linked antibody screening system.

机构信息

Laboratory for Integrative Genomics, RIKEN Center for Integrative Medical Sciences, Yokohama, Japan.

Laboratory for Lymphocyte Differentiation, RIKEN Center for Integrative Medical Sciences, Yokohama, Japan.

出版信息

Elife. 2024 Nov 19;13:RP95346. doi: 10.7554/eLife.95346.

DOI:10.7554/eLife.95346
PMID:39558690
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11575895/
Abstract

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.

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方法外，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限制：（1）筛选仅在 Ig 基因克隆和重组抗体生产之后开始，以及（2）抗体由配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，我们通过建立基于 Golden Gate 的双表达载体和膜结合抗体的体内表达，介绍了一种快速筛选重组单克隆抗体的方法。使用该系统，我们在 7 天内从免疫小鼠中快速分离出具有高亲和力的流感交叉反应性抗体。该系统特别适用于分离治疗性或诊断性抗体，例如在预期的大流行期间。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/ec172b09beaa/elife-95346-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/f7598f5e232b/elife-95346-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/908160851abc/elife-95346-fig1-figsupp1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/bb32f2b43292/elife-95346-fig1-figsupp2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/63a4e5d2590a/elife-95346-fig1-figsupp3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/985f3ab2c6c4/elife-95346-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/cf698d133518/elife-95346-fig2-figsupp1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/f2c8a8773be1/elife-95346-fig2-figsupp2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/bdd0cb4960e8/elife-95346-fig2-figsupp3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/ec172b09beaa/elife-95346-fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/f7598f5e232b/elife-95346-fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/908160851abc/elife-95346-fig1-figsupp1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/bb32f2b43292/elife-95346-fig1-figsupp2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/63a4e5d2590a/elife-95346-fig1-figsupp3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/985f3ab2c6c4/elife-95346-fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/cf698d133518/elife-95346-fig2-figsupp1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/f2c8a8773be1/elife-95346-fig2-figsupp2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/bdd0cb4960e8/elife-95346-fig2-figsupp3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0932/11575895/ec172b09beaa/elife-95346-fig3.jpg

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High-Throughput Mapping of B Cell Receptor Sequences to Antigen Specificity.高通量 B 细胞受体序列到抗原特异性的映射。
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