Volume 10 Issue 1
Feb.  2024
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Shenglan Zhang, Zhiran Fan, Jianfeng Liu. Generation and characterization of nanobodies targeting GPCR. Biophysics Reports, 2024, 10(1): 22-30. doi: 10.52601/bpr.2023.230026
Citation: Shenglan Zhang, Zhiran Fan, Jianfeng Liu. Generation and characterization of nanobodies targeting GPCR. Biophysics Reports, 2024, 10(1): 22-30. doi: 10.52601/bpr.2023.230026

Generation and characterization of nanobodies targeting GPCR

doi: 10.52601/bpr.2023.230026
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  • Corresponding author: jfliu@mail.hust.edu.cn (J. Liu)
  • Received Date: 25 October 2023
  • Accepted Date: 15 December 2023
  • Available Online: 26 February 2024
  • Publish Date: 29 February 2024
  • 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.

  • 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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