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Nanobody Production and Purification

FZ Fang Zheng
JZ Jinhong Zhou
ZO Zhenlin Ouyang
JZ Jiaxin Zhang
XW Xinyi Wang
SM Serge Muyldermans
JG Jo Van Ginderachter
ND Nick Devoogdt
YW Yurong Wen
SS Steve Schoonooghe
GR Geert Raes
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The Clec4F-specific nanobody gene sequences were inserted into the pHEN6c plasmid and transfected into E. coli WK6 host cells. Hexahistidine-tagged (His-tag) Clec4F nanobodies were extracted from E. coli WK6 periplasmic space by osmotic shock. All the extracted proteins were expressed in 400 mL medium and purified by immobilized metal affinity chromatography (IMAC) using the Ni-NTA column and the buffer was exchanged from 0.5 M imidazole elution buffer to PBS by dialysis. The purity and size of the nanobody was detected by SDS-PAGE and Coomassie brilliant blue staining.

Copyright and License information:  ©2021 Zheng, Zhou, Ouyang, Zhang, Wang, Muyldermans, Van Ginderachter, Devoogdt, Wen, Schoonooghe and Raes.
This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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