Fast Epitope Mapping for the Anti‐MUC1 Monoclonal Antibody by Combining a One‐Bead‐One‐Glycopeptide Library and a Microarray Platform |
| |
| Authors: | Dr. Fayna Garcia‐Martin Dr. Takahiko Matsushita Dr. Hiroshi Hinou Prof. Dr. Shin‐Ichiro Nishimura |
| |
| Affiliation: | 1. Faculty of Advanced Life Science and Graduate School of Life Science, Hokkaido University, N21, W11, Kita‐ku, Sapporo 001‐0021 (Japan), Fax: (+81)?11‐706‐9042;2. Medicinal Chemistry Pharmaceuticals, Co. Ltd. N21, W12, Kita‐ku, Sapporo 001‐0021 (Japan) |
| |
| Abstract: | 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 O‐glycan 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. |
| |
| Keywords: | antibodies combinatorial chemistry glycopeptides microarrays solid‐phase synthesis |
|
|
