Generation and characterization of nanobodies targeting GPCR  

作　　者：Shenglan Zhang Zhiran Fan Jianfeng Liu 

机构地区：[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 Sci-ence and Technology,Wuhan 430074,China

出　　处：《Biophysics Reports》2024年第1期22-30,共9页生物物理学报（英文版）

基　　金：financially supported by the Key Technologies R&D Program of Guangdong Province(2010A080813001);GuangZhou Basic and Applied Basic Research Foundation(SL2022A04J00399).

摘　　要：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 anti-bodies,HcAbs)antibodies,are small antibody fragments derived from camelids that have gained signif-icant 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 sur-face.We developed a universal protocol for making nanobodies targeting GPCRs using the cell mem-brane 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 mono-clonal nanobodies were recombinantly expressed in Escherichia coli(E.coli)and purified to character-ize their binding potency.

关 键 词：NANOBODY Single domain antibody Heavy chain-only antibody GPCR G protein-coupled receptor 

分 类 号：R392-33[医药卫生—免疫学]