The screening and identification of internalized nanobody against EpCAM / 药学学报
Acta Pharmaceutica Sinica
; (12): 2405-2413, 2020.
Article
de Zh
| WPRIM
| ID: wpr-829388
Bibliothèque responsable:
WPRO
ABSTRACT
Epithelial cell adhesion molecule (EpCAM) is a popular target for cancer therapy. In this research, 3 nanobodies with high specificity and endocytosis activity against EpCAM were developed, which provides a basis for the study of immunotoxin based on EpCAM. In our preliminary experiments, we have immunized a camel with EpCAM-Fc antigen and constructed a high-quality phage display library. Seventeen nanobodies with different complementarity determining region (CDR) 3 sequences have been screened after 3 rounds of biopanning by phage display technology. The animal procedures were approved by the Institutional Animal Care and Use Committee (IACUC) of Fudan University School of Pharmacy. After purification, 7 nanobodies showed high cell binding activity by fluorescent activated cell sorting (FACS) identification. Furthermore, 3 nanobodies presented high endocytosis activity based on FACS and laser confocal microscopy, which also showed high affinity to EpCAM measured by ForteBio. According to this study, we aimed to provide a novel alternative approach to the EpCAM-targeted therapy and to provide guidance for the study of nanobody based immunotoxins for other targets.
Texte intégral:
1
Indice:
WPRIM
Type d'étude:
Diagnostic_studies
/
Guideline
/
Screening_studies
langue:
Zh
Texte intégral:
Acta Pharmaceutica Sinica
Année:
2020
Type:
Article