| RRC ID |
82087
|
| Author |
Lin N, Miyamoto K, Ogawara T, Sakurai S, Kizaka-Kondoh S, Kadonosono T.
|
| Title |
Epitope binning for multiple antibodies simultaneously using mammalian cell display and DNA sequencing.
|
| Journal |
Commun Biol
|
| Abstract |
Epitope binning, an approach for grouping antibodies based on epitope similarities, is a critical step in antibody drug discovery. However, conventional methods are complex, involving individual antibody production. Here, we established Epitope Binning-seq, an epitope binning platform for simultaneously analyzing multiple antibodies. In this system, epitope similarity between the query antibodies (qAbs) displayed on antigen-expressing cells and a fluorescently labeled reference antibody (rAb) targeting a desired epitope is analyzed by flow cytometry. The qAbs with epitope similar to the rAb can be identified by next-generation sequencing analysis of fluorescence-negative cells. Sensitivity and reliability of this system are confirmed using rAbs, pertuzumab and trastuzumab, which target human epidermal growth factor receptor 2. Epitope Binning-seq enables simultaneous epitope evaluation of 14 qAbs at various abundances in libraries, grouping them into respective epitope bins. This versatile platform is applicable to diverse antibodies and antigens, potentially expediting the identification of clinically useful antibodies.
|
| Volume |
7(1)
|
| Pages |
652
|
| Published |
2024-5-28
|
| DOI |
10.1038/s42003-024-06363-7
|
| PII |
10.1038/s42003-024-06363-7
|
| PMID |
38806676
|
| PMC |
PMC11133372
|
| MeSH |
Animals
Antibodies / genetics
Antibodies / immunology
Antibodies, Monoclonal, Humanized / immunology
Epitope Mapping / methods
Epitopes* / immunology
Flow Cytometry / methods
High-Throughput Nucleotide Sequencing / methods
Humans
Receptor, ErbB-2 / genetics
Receptor, ErbB-2 / immunology
Sequence Analysis, DNA / methods
Trastuzumab / immunology
|
| IF |
4.165
|
| Resource |
| Human and Animal Cells |
293(RCB1637)
293T(RCB2202) |
