A perspective toward mass spectrometry-based de novo sequencing of endogenous antibodies.
A perspective toward mass spectrometry-based de novo sequencing of endogenous antibodies.
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DOI：
10.1080/19420862.2022.2079449
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发表时间：
2022-01
期刊：
影响因子：
5.3
通讯作者：
Heck, Albert J. R.
中科院分区：
文献类型：
--
作者：
de Graaf, Sebastiaan C.;Hoek, Max;Tamara, Sem;Heck, Albert J. R.
治疗性和内源性体液抗体表征的关键步骤是鉴定氨基酸序列。到目前为止，这项任务主要是通过在核苷酸水平上对B细胞受体（BCR）库进行测序来解决的。质谱（MS）已经成为一种替代工具，用于直接在最相关的蛋白质水平上获得序列信息。虽然现在已经建立了几种MS方法，但重组和内源性抗体的分析面临着一系列特定的挑战，需要超越传统蛋白质组学工作流程的方法。在这里，我们回顾了重组以及内源性体液抗体的MS测序的挑战，并概述了试图克服这些障碍的最先进的方法。我们强调最近的例子，并讨论仍然存在的挑战。我们预见了这些方法的巨大前景，使得通过基于MS的技术进行从头抗体测序和发现变得可行，即使对于来自内源性来源的复杂临床样品，如血清和其他液体活检。
A key step in therapeutic and endogenous humoral antibody characterization is identifying the amino acid sequence. So far, this task has been mainly tackled through sequencing of B-cell receptor (BCR) repertoires at the nucleotide level. Mass spectrometry (MS) has emerged as an alternative tool for obtaining sequence information directly at the – most relevant – protein level. Although several MS methods are now well established, analysis of recombinant and endogenous antibodies comes with a specific set of challenges, requiring approaches beyond the conventional proteomics workflows. Here, we review the challenges in MS-based sequencing of both recombinant as well as endogenous humoral antibodies and outline state-of-the-art methods attempting to overcome these obstacles. We highlight recent examples and discuss remaining challenges. We foresee a great future for these approaches making de novo antibody sequencing and discovery by MS-based techniques feasible, even for complex clinical samples from endogenous sources such as serum and other liquid biopsies.
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