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高通量克隆、表达、纯化和双特异性抗体评估的方案。

Protocol for high-throughput cloning, expression, purification, and evaluation of bispecific antibodies.

机构信息

Department of Therapeutics Discovery, Amgen Research, Amgen Inc., Thousand Oaks, CA 91320, USA.

Department of Therapeutics Discovery, Amgen Research, Amgen Inc., San Francisco, CA 94080, USA.

出版信息

STAR Protoc. 2022 May 27;3(2):101428. doi: 10.1016/j.xpro.2022.101428. eCollection 2022 Jun 17.

DOI:10.1016/j.xpro.2022.101428
PMID:35664258
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9157557/
Abstract

Bispecific antibodies are a powerful new class of therapeutics, but their development often requires enormous amounts of time and resources. Here, we describe a high-throughput protocol for cloning, expressing, purifying, and evaluating bispecific antibodies. This protocol enables the rapid screening of large panels of bispecific molecules to identify top candidates for further development. For complete details on the use and execution of this protocol, please refer to Estes et al. (2021).

摘要

双特异性抗体是一类强大的新型治疗药物，但它们的开发通常需要大量的时间和资源。在这里，我们描述了一种用于克隆、表达、纯化和评估双特异性抗体的高通量方案。该方案能够快速筛选大量双特异性分子，以鉴定进一步开发的顶级候选物。有关该方案使用和执行的完整详细信息，请参阅 Estes 等人。(2021)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e89e/9157557/97fc27bbf888/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e89e/9157557/a21ea14bc258/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e89e/9157557/06d2c77549bc/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e89e/9157557/c260e3c23af2/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e89e/9157557/9736f72dc77b/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e89e/9157557/97fc27bbf888/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e89e/9157557/a21ea14bc258/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e89e/9157557/06d2c77549bc/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e89e/9157557/c260e3c23af2/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e89e/9157557/9736f72dc77b/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e89e/9157557/97fc27bbf888/gr4.jpg

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Rational selection of building blocks for the assembly of bispecific antibodies.理性选择构建双特异性抗体的构建块。
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High-throughput antibody screening from complex matrices using intact protein electrospray mass spectrometry.利用完整蛋白质电喷雾质谱对复杂基质中的抗体进行高通量筛选。
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Nat Rev Drug Discov. 2019 Aug;18(8):585-608. doi: 10.1038/s41573-019-0028-1.
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Unraveling the mysteries of modern size exclusion chromatography - the way to achieve confident characterization of therapeutic proteins.揭开现代尺寸排阻色谱法的奥秘 - 实现治疗性蛋白质可靠表征的方法。
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