2014年11月
Fast Epitope Mapping for the Anti-MUC1 Monoclonal Antibody by Combining a One-Bead-One-Glycopeptide Library and a Microarray Platform
CHEMISTRY-A EUROPEAN JOURNAL
- ,
- ,
- ,
- 巻
- 20
- 号
- 48
- 開始ページ
- 15891
- 終了ページ
- 15902
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1002/chem.201403239
- 出版者・発行元
- WILEY-V C H VERLAG GMBH
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.
- リンク情報
- ID情報
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- DOI : 10.1002/chem.201403239
- ISSN : 0947-6539
- eISSN : 1521-3765
- PubMed ID : 25303614
- Web of Science ID : WOS:000345234900032