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The screening and identification of internalized nanobody against EpCAM
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Zong-shu XIAN1, Guang-hui LI2, Jun-wei GAI2, Min ZHU2, Lin-lin MA3, Dian-wen JU1, *, Ya-kun WAN2, *
Acta Pharmaceutica Sinica | 2020, 55(10) : 2405 - 2413
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Acta Pharmaceutica Sinica | 2020, 55(10): 2405-2413
• Original Articles •
The screening and identification of internalized nanobody against EpCAM
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Zong-shu XIAN1, Guang-hui LI2, Jun-wei GAI2, Min ZHU2, Lin-lin MA3, Dian-wen JU1, *, Ya-kun WAN2, *
Affiliations
  • 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
Published: 2020-10-12 doi: 10.16438/j.0513-4870.2020-1359
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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.

epithelial cell adhesion molecule  /  nanobody  /  phage display technology  /  immune library  /  endocytosis
Zong-shu XIAN, Guang-hui LI, Jun-wei GAI, Min ZHU, Lin-lin MA, Dian-wen JU, Ya-kun WAN. The screening and identification of internalized nanobody against EpCAM[J]. Acta Pharmaceutica Sinica, 2020 , 55 (10) : 2405 -2413 . DOI: 10.16438/j.0513-4870.2020-1359
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Article Info
doi: 10.16438/j.0513-4870.2020-1359
  • Receive Date：2020-08-18
  • Online Date：2026-01-23
  • Published：2020-10-12
Article Data
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History
  • Received：2020-08-18
  • Revised：2020-09-09
Funding
Affiliations
    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
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表12种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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