The screening and identification of internalized nanobody against EpCAM
10.16438/j.0513-4870.2020-1359
- VernacularTitle:内吞型EpCAM纳米抗体的筛选及鉴定
- Author:
Zong-shu XIAN
1
;
Guang-hui LI
2
;
Jun-wei GAI
2
;
Min ZHU
2
;
Lin-lin MA
3
;
Dian-wen JU
1
;
Ya-kun WAN
2
Author Information
1. School of Pharmacy, Fudan University, Shanghai 201203, China
2. Shanghai Novamab Biopharmaceuticals Co., Ltd., Shanghai 201203, China
3. Shanghai Key Laboratory of Molecular Imaging, Shanghai University of Medicine and Health Sciences, Shanghai 201318, China
- Publication Type:Research Article
- Keywords:
epithelial cell adhesion molecule;
nanobody;
phage display technology;
immune library;
endocytosis
- From:
Acta Pharmaceutica Sinica
2020;55(10):2405-2413
- CountryChina
- Language:Chinese
-
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.
