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首页> 外文期刊>Chemistry: A European journal >Fast Epitope Mapping for the Anti-MUC1 Monoclonal Antibody by Combining a One-Bead-One-Glycopeptide Library and a Microarray Platform
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Fast Epitope Mapping for the Anti-MUC1 Monoclonal Antibody by Combining a One-Bead-One-Glycopeptide Library and a Microarray Platform

机译：通过组合一个珠子一个糖肽库和一个微阵列平台的抗MUC1单克隆抗体的快速表位作图。

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摘要

Anti-MUC1 monoclonal antibodies (mAbs) are powerful tools that can be used to recognize cancer-related MUC1 molecules, the O-glycosylation status of which is believed to affect binding affinity. We demonstrate the feasibility of using a rapid screening methodology to elucidate those effects. The approach involves i) " one-bead-one-compound"- based preparation of bilayer resins carrying glycopeptides on the shell and mass-tag tripeptides coding Oglycan patterns in the core, ii) on-resin screening with an anti-MUC1 mAb, iii) separating positive resins by utilizing secondary antibody conjugation with magnetic beads, and (iv) decoding the mass-tag that is detached from the positive resins pool by using mass spectrometric analysis. We tested a small library consisting of 27 MUC1 glycopeptides with different O-glycosylations against anti-MUC1 mAb clone VU-3C6. Qualitative mass-tag analysis showed that increasing the number of glycans leads to an increase in the binding affinity. Six glycopeptides selected from the library were validated by using a microarray-based assay. Our screening provides valuable information on O-glycosylations of epitopes leading to high affinity with mAb.
机译：抗MUC1单克隆抗体（mAbs）是强大的工具，可用于识别与癌症相关的MUC1分子，其O-糖基化状态被认为会影响结合亲和力。我们证明了使用快速筛查方法阐明这些影响的可行性。该方法涉及以下步骤：i）基于“单珠一化合物”的双层树脂制备，在壳上带有糖肽和在核心上编码糖链模式的质量标签三肽，ii）用抗MUC1 mAb在树脂上进行筛选， iii）通过利用与磁珠结合的二抗来分离阳性树脂，以及（iv）通过质谱分析对从阳性树脂库中分离的质量标签进行解码。我们测试了一个由27个MUC1糖肽组成的小文库，它们针对抗MUC1 mAb克隆VU-3C6具有不同的O-糖基化。定性质量标签分析表明，聚糖数量的增加导致结合亲和力的增加。使用基于微阵列的测定法验证了从文库中选出的六个糖肽。我们的筛选提供了有关表位的O-糖基化的有价值信息，这些信息可导致与mAb的高度亲和力。

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