Toxins | |
Uniform Orientation of Biotinylated Nanobody as an Affinity Binder for Detection of |
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Min Li2  Min Zhu2  Cunzheng Zhang1  Xianjin Liu1  Yakun Wan2  | |
[1] Institute of Food Safety, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China; E-Mail:;The Key Laboratory of Developmental Genes and Human Disease, Ministry of Education, Institute of Life Sciences, Sipailou NO. 2, Southeast University, Nanjing 210096, China; E-Mails: | |
关键词: nanobody; Cry1Ac toxin; streptavidin; DAS-ELISA; | |
DOI : 10.3390/toxins6123208 | |
来源: mdpi | |
【 摘 要 】
Nanobodies are the smallest natural fragments with useful properties such as high affinity, distinct paratope and high stability, which make them an ideal tool for detecting target antigens. In this study, we generated and characterized nanobodies against the Cry1Ac toxin and applied them in a biotin-streptavidin based double antibodies (nanobodies) sandwich-ELISA (DAS-ELISA) assay. After immunizing a camel with soluble Cry1Ac toxin, a phage displayed library was constructed to generate Nbs against the Cry1Ac toxin. Through successive rounds of affinity bio-panning, four nanobodies with greatest diversity in CDR3 sequences were obtained. After affinity determination and conjugating to HRP, two nanobodies with high affinity which can recognize different epitopes of the same antigen (Cry1Ac) were selected as capture antibody (Nb61) and detection antibody (Nb44). The capture antibody (Nb61) was biotinylated
【 授权许可】
CC BY
© 2014 by the authors; licensee MDPI, Basel, Switzerland.
【 预 览 】
Files | Size | Format | View |
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RO202003190018897ZK.pdf | 1967KB | download |