This is the first report on the screening,expression,and recognition mechanism analysis of single-chain fragment variable(scFv)against phenylethanolamine A(PEAA),a newly emergedβ-adrenergic agonist illegally used as ...This is the first report on the screening,expression,and recognition mechanism analysis of single-chain fragment variable(scFv)against phenylethanolamine A(PEAA),a newly emergedβ-adrenergic agonist illegally used as a feed additive for growth promotion.The PEAA-specific scFv scFv,called scFv-32,was screened from hybridoma cell lines by phage display and was found to be optimally expressed in the E.coli system.The ic-ELISA results revealed an IC_(50)value of 10.34μg/L for scFv-32 and no cross-reactivity with otherβ-adrenergic agonists.Homology modeling and molecular docking revealed the key binding sites VAL178,TYP228,and ASP229.One hydrogen bond,two pisigma bonds,and one pi-pi bond maintain the formation of the antibody‒drug complex.Alanine scanning mutagenesis of the three predicted key binding sites showed that the mutants completely lost their recognition activity,which confirmed the accuracy of the theoretical analysis.These results are valuable for the preparation of scFvs and the analysis of the molecular recognition mechanism of antigen-antibodies.展开更多
The discovery of G-protein coupled receptor(GPCR)accessory proteins has fundamentally redefined the pharmacological concept of GPCR signaling,demonstrating a more complex molecular basis for receptor specificity on th...The discovery of G-protein coupled receptor(GPCR)accessory proteins has fundamentally redefined the pharmacological concept of GPCR signaling,demonstrating a more complex molecular basis for receptor specificity on the plasma membrane and impressionable downstream intracellular cascades.GPCR accessory proteins not only contribute to the proper folding and trafficking of receptors but also exhibit selectable receptor preferences.展开更多
As a powerful cell-based therapeutic approach,CAR-T therapy was originally designed for treating acquired immunodeficiency syndrome(AIDS)(Baker et al.,2023),but had been strikingly successful in curing hematologic mal...As a powerful cell-based therapeutic approach,CAR-T therapy was originally designed for treating acquired immunodeficiency syndrome(AIDS)(Baker et al.,2023),but had been strikingly successful in curing hematologic malignancies and multiple solid tumors.Numerous evidence has expanded the medical application of CAR-T therapy for the treatment of many other human diseases beyond cancer.In this article,we discuss the principle of CAR-T and enumerate the current application and limitation in oncology.Finally,we provide a comprehensive perspective of current advance and future directions of CAR-T in treating multiple non-tumoral diseases.展开更多
基金the National Natural Science Foundation of China(32072920)the Fundamental Research Funds for the Central Universities(2662022DKPY007)the HZAU-AGIS Cooperation Fund(SZYJY2022024).
文摘This is the first report on the screening,expression,and recognition mechanism analysis of single-chain fragment variable(scFv)against phenylethanolamine A(PEAA),a newly emergedβ-adrenergic agonist illegally used as a feed additive for growth promotion.The PEAA-specific scFv scFv,called scFv-32,was screened from hybridoma cell lines by phage display and was found to be optimally expressed in the E.coli system.The ic-ELISA results revealed an IC_(50)value of 10.34μg/L for scFv-32 and no cross-reactivity with otherβ-adrenergic agonists.Homology modeling and molecular docking revealed the key binding sites VAL178,TYP228,and ASP229.One hydrogen bond,two pisigma bonds,and one pi-pi bond maintain the formation of the antibody‒drug complex.Alanine scanning mutagenesis of the three predicted key binding sites showed that the mutants completely lost their recognition activity,which confirmed the accuracy of the theoretical analysis.These results are valuable for the preparation of scFvs and the analysis of the molecular recognition mechanism of antigen-antibodies.
基金supported by the National Natural Science Foundation of China(Grant No.32271165)Shanghai Clinical Research Center of Plastic and Reconstructive Surgery supported by the Science and Technology Commission of Shanghai Municipality(Grant No.22MC1940300)+2 种基金the Innovative Research Team of High-level Local Universities in Shanghai(Grant No.SHSMU-ZDCX20210400)the Key Laboratory Program of the Education Commission of Shanghai Municipality(No.ZDSYS14005)China Postdoctoral Science Foundation(Grant No.2022M722127).
文摘The discovery of G-protein coupled receptor(GPCR)accessory proteins has fundamentally redefined the pharmacological concept of GPCR signaling,demonstrating a more complex molecular basis for receptor specificity on the plasma membrane and impressionable downstream intracellular cascades.GPCR accessory proteins not only contribute to the proper folding and trafficking of receptors but also exhibit selectable receptor preferences.
基金supported by the National Key Research and Development Program of China(Grant no.2019YFA0111400,2022YFE0200100)the National Natural Science Foundation of China(Grant no.32271165,82070144,82270155)the Interdisciplinary Project of Central Universities(Grant no.2022-2-ZD-02).
文摘As a powerful cell-based therapeutic approach,CAR-T therapy was originally designed for treating acquired immunodeficiency syndrome(AIDS)(Baker et al.,2023),but had been strikingly successful in curing hematologic malignancies and multiple solid tumors.Numerous evidence has expanded the medical application of CAR-T therapy for the treatment of many other human diseases beyond cancer.In this article,we discuss the principle of CAR-T and enumerate the current application and limitation in oncology.Finally,we provide a comprehensive perspective of current advance and future directions of CAR-T in treating multiple non-tumoral diseases.
