首页|Generation and characterization of nanobodies targeting GPCR
Generation and characterization of nanobodies targeting GPCR
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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.
Bio-land Laboratory,Guangzhou Regenerative Medicine and Health Guangdong Laboratory,Guangzhou 510005,China
Bioland Laboratory(Guangzhou Regenerative Medicine and Health Guangdong Laboratory),Guangzhou 510005,China
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
Key Technologies Research and Development Program of Guangdong ProvinceGuangzhou Basic and Applied Basic Research Foundation
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