| RRC ID |
68946
|
| 著者 |
See K, Kadonosono T, Miyamoto K, Tsubaki T, Ota Y, Katsumi M, Ryo S, Aida K, Minegishi M, Isozaki T, Kuchimaru T, Kizaka-Kondoh S.
|
| タイトル |
Antibody-guided design and identification of CD25-binding small antibody mimetics using mammalian cell surface display.
|
| ジャーナル |
Sci Rep
|
| Abstract |
Small antibody mimetics that contain high-affinity target-binding peptides can be lower cost alternatives to monoclonal antibodies (mAbs). We have recently developed a method to create small antibody mimetics called FLuctuation-regulated Affinity Proteins (FLAPs), which consist of a small protein scaffold with a structurally immobilized target-binding peptide. In this study, to further develop this method, we established a novel screening system for FLAPs called monoclonal antibody-guided peptide identification and engineering (MAGPIE), in which a mAb guides selection in two manners. First, antibody-guided design allows construction of a peptide library that is relatively small in size, but sufficient to identify high-affinity binders in a single selection round. Second, in antibody-guided screening, the fluorescently labeled mAb is used to select mammalian cells that display FLAP candidates with high affinity for the target using fluorescence-activated cell sorting. We demonstrate the reliability and efficacy of MAGPIE using daclizumab, a mAb against human interleukin-2 receptor alpha chain (CD25). Three FLAPs identified by MAGPIE bound CD25 with dissociation constants of approximately 30 nM as measured by biolayer interferometry without undergoing affinity maturation. MAGPIE can be broadly adapted to any mAb to develop small antibody mimetics.
|
| 巻・号 |
11(1)
|
| ページ |
22098
|
| 公開日 |
2021-11-11
|
| DOI |
10.1038/s41598-021-01603-w
|
| PII |
10.1038/s41598-021-01603-w
|
| PMID |
34764369
|
| PMC |
PMC8585965
|
| MeSH |
Amino Acid Sequence
Animals
Antibodies, Monoclonal / immunology*
Antibody Affinity / immunology
Cell Line
Cell Line, Tumor
Cell Surface Display Techniques / methods*
Flow Cytometry / methods
HEK293 Cells
HeLa Cells
Humans
Interleukin-2 Receptor alpha Subunit / immunology*
K562 Cells
Mammals / immunology*
Peptide Library
Protein Binding / immunology*
|
| IF |
3.998
|
| リソース情報 |
| ヒト・動物細胞 |
293(RCB1637)
HeLa(RCB0007) |
