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. 2021 Dec 8;9:792111. doi: 10.3389/fbioe.2021.792111

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Copyright © 2021 Wei, Wang, Xiang, Jiang, Gong, Su, Al Azad, Dong, Feng, Wu, Chan, Yang and Shi.

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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Validation of GFP-specific vNARs as intrabodies and bivalent constructs. (A) Schematic for vNARs as intrabodies in human cells. (B) Expression of vNARs in HEK293T cells. (C) Native GFP-vNAR complexes were pulled down from cell lysates. (D) Design of bivalent vNARs. (E) Mapping of binding epitopes. (F) SDS-PAGE of purified bivalent vNARs. (G) SPR sensorgram of BsG3-BsG98 binding with GFP. (H) Binding affinities of four bivalent vNARs to GFP. The values represent mean ± standard error. (I) Schematic for the identification of high-affinity binders from bamboo shark vNAR library and their applications.