Generation and characterization of nanobodies targeting GPCR

Shenglan Zhang; Zhiran Fan; Jianfeng Liu

doi:10.52601/bpr.2023.230026

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Shenglan Zhang, Zhiran Fan, Jianfeng Liu. Generation and characterization of nanobodies targeting GPCR. Biophysics Reports. doi: 10.52601/bpr.2023.230026

Citation:

Shenglan Zhang, Zhiran Fan, Jianfeng Liu. Generation and characterization of nanobodies targeting GPCR. Biophysics Reports. doi: 10.52601/bpr.2023.230026

Shenglan Zhang, Zhiran Fan, Jianfeng Liu. Generation and characterization of nanobodies targeting GPCR. Biophysics Reports. doi: 10.52601/bpr.2023.230026

Citation:

Shenglan Zhang, Zhiran Fan, Jianfeng Liu. Generation and characterization of nanobodies targeting GPCR. Biophysics Reports. doi: 10.52601/bpr.2023.230026

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Generation and characterization of nanobodies targeting GPCR

doi: 10.52601/bpr.2023.230026

Shenglan Zhang^{1, S},
Zhiran Fan^{1, S},
Jianfeng Liu^{1, 2
,
,}

1.
Bioland Laboratory (Guangzhou Regenerative Medicine and Health Guangdong Laboratory), Guangzhou 510005, China
2.
Cellular Signaling laboratory, International Research Center for Sensory Biology and Technology of MOST, Key Laboratory of Molecular Biophysics of MOE, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China

More Information

Corresponding author: jfliu@mail.hust.edu.cn (J. Liu)
Received Date: 25 October 2023
Accepted Date: 15 December 2023

Available Online: 26 February 2024

Abstract

Abstract

G protein-coupled receptors (GPCRs) are a large family of cell membrane proteins that are important targets for drug discovery. Nanobodies, also known as VHH (variable domains of heavy chain-only antibodies, HcAbs) antibodies, are small antibody fragments derived from camelids that have gained significant attention as potential therapeutics for targeting GPCRs due to their advantages over conventional antibodies. However, there are challenges in developing nanobodies targeting GPCRs, among which epitope accessibility is the most significant because the cell membrane partially shields the GPCR surface. We developed a universal protocol for making nanobodies targeting GPCRs using the cell membrane extract of GPCR-overexpressing HEK293 cells as the llama/alpaca immunization antigen. We constructed an immune VHH library and identified nanobodies by phage display bio-panning. The monoclonal nanobodies were recombinantly expressed in Escherichia coli (E. coli) and purified to characterize their binding potency.
- Nanobody,
- Single domain antibody,
- Heavy chain-only antibody,
- GPCR,
- G protein-coupled receptor

Shenglan Zhang, Zhiran Fan and Jianfeng Liu declare that they have no conflict of insterest.
All institutional and national guidelines for the care and use of laboratory animals were followed.
Shenglan Zhang and Zhiran Fan contributed equally to this work.

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References(26)

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