目的构建可以同时靶向免疫检查点分子CD96和PD1(programmed cell death protein 1)的双特异性抗体,为肿瘤免疫治疗提供一种新的思路。方法构建表达抗CD96和PD1的ScFv-Fc结构的质粒,使用CHO真核表达系统生产抗体。经过Protein A纯化后,使...目的构建可以同时靶向免疫检查点分子CD96和PD1(programmed cell death protein 1)的双特异性抗体,为肿瘤免疫治疗提供一种新的思路。方法构建表达抗CD96和PD1的ScFv-Fc结构的质粒,使用CHO真核表达系统生产抗体。经过Protein A纯化后,使用SDS-PAGE和分子排阻色谱鉴定抗体。外源构建CHO-PD1和293T-CD962种靶细胞,流式细胞术检测CD96和PD1的ScFv-Fc抗体与上述靶细胞以及T细胞上抗原的结合能力,并采用Fortebio分子互作仪进行抗原抗体结合解离动力学验证。分别采用Knobs-into-holes(KIH)突变策略构建ScFv-KIH双特异性抗体结构,以及类DVD-Ig(Dual variable domain immunoglobulin)分子结构,构建ScFv串联型(ScFv)2-Fc结构的双特异性抗体,并分别对表达的2种双特异性抗体进行了功能验证。结果抗CD96和PD1的ScFv-Fc抗体可特异性识别靶细胞和T细胞上的靶抗原,构建生产的2种结构的双特异性抗体均能正常表达,且能同时靶向CD96和PD12种抗原。结论ScFv-KIH和(ScFv)2-Fc型的双特异性抗体表达成功,并可同时识别CD96和PD1抗原。展开更多
Bispecific antibodies(bsAbs)refer to a large family of molecules that recognize two different epitopes or antigens.Although a series of challenges,especially immunogenicity and chain mispairing issues,once hindered th...Bispecific antibodies(bsAbs)refer to a large family of molecules that recognize two different epitopes or antigens.Although a series of challenges,especially immunogenicity and chain mispairing issues,once hindered the development of bsAbs,they have been gradually overcome with the help of rapidly developing technologies in the past 5 decades.In the meantime,an increasing number of bsAb platforms have been designed to satisfy different clinical demands.Currently,numerous preclinical and clinical trials are underway,portraying a promising future for bsAb-based cancer treatment.Nevertheless,bsAb drugs still face enormous challenges in their application as cancer therapeutics,including tumor heterogeneity and mutational burden,intractable tumor microenvironment(TME),insufficient costimulatory signals to activate T cells,the necessity for continuous injection,fatal systemic side effects,and off-target toxicities to adjacent normal cells.Therefore,we provide several strategies as solutions to these issues,which comprise generating multispecific bsAbs,discovering neoantigens,combining bsAbs with other anticancer therapies,exploiting natural killer(NK)-cell-based bsAbs and producing bsAbs in situ.In this review,we mainly discuss previous and current challenges in bsAb development and underscore corresponding strategies,with a brief introduction of several typical bsAb formats.展开更多
T cell-redirecting bispecific antibodies are specifically designed to bind to tumor-associated antigens,thereby engaging with CD3 on the T cell receptor.This linkage between tumor cells and T cells actively triggers T...T cell-redirecting bispecific antibodies are specifically designed to bind to tumor-associated antigens,thereby engaging with CD3 on the T cell receptor.This linkage between tumor cells and T cells actively triggers T cell activation and initiates targeted killing of the identified tumor cells.These antibodies have emerged as one of the most promising avenues within tumor immunotherapy.However,despite success in treating hematological malignancies,significant advancements in solid tumors have yet to be explored.In this review,we aim to address the critical challenges associated with T cellredirecting bispecific antibodies and explore novel strategies to overcome these obstacles,with the ultimate goal of expanding the application of this therapy to include solid tumors.展开更多
文摘目的构建可以同时靶向免疫检查点分子CD96和PD1(programmed cell death protein 1)的双特异性抗体,为肿瘤免疫治疗提供一种新的思路。方法构建表达抗CD96和PD1的ScFv-Fc结构的质粒,使用CHO真核表达系统生产抗体。经过Protein A纯化后,使用SDS-PAGE和分子排阻色谱鉴定抗体。外源构建CHO-PD1和293T-CD962种靶细胞,流式细胞术检测CD96和PD1的ScFv-Fc抗体与上述靶细胞以及T细胞上抗原的结合能力,并采用Fortebio分子互作仪进行抗原抗体结合解离动力学验证。分别采用Knobs-into-holes(KIH)突变策略构建ScFv-KIH双特异性抗体结构,以及类DVD-Ig(Dual variable domain immunoglobulin)分子结构,构建ScFv串联型(ScFv)2-Fc结构的双特异性抗体,并分别对表达的2种双特异性抗体进行了功能验证。结果抗CD96和PD1的ScFv-Fc抗体可特异性识别靶细胞和T细胞上的靶抗原,构建生产的2种结构的双特异性抗体均能正常表达,且能同时靶向CD96和PD12种抗原。结论ScFv-KIH和(ScFv)2-Fc型的双特异性抗体表达成功,并可同时识别CD96和PD1抗原。
文摘Bispecific antibodies(bsAbs)refer to a large family of molecules that recognize two different epitopes or antigens.Although a series of challenges,especially immunogenicity and chain mispairing issues,once hindered the development of bsAbs,they have been gradually overcome with the help of rapidly developing technologies in the past 5 decades.In the meantime,an increasing number of bsAb platforms have been designed to satisfy different clinical demands.Currently,numerous preclinical and clinical trials are underway,portraying a promising future for bsAb-based cancer treatment.Nevertheless,bsAb drugs still face enormous challenges in their application as cancer therapeutics,including tumor heterogeneity and mutational burden,intractable tumor microenvironment(TME),insufficient costimulatory signals to activate T cells,the necessity for continuous injection,fatal systemic side effects,and off-target toxicities to adjacent normal cells.Therefore,we provide several strategies as solutions to these issues,which comprise generating multispecific bsAbs,discovering neoantigens,combining bsAbs with other anticancer therapies,exploiting natural killer(NK)-cell-based bsAbs and producing bsAbs in situ.In this review,we mainly discuss previous and current challenges in bsAb development and underscore corresponding strategies,with a brief introduction of several typical bsAb formats.
基金supported by the National Natural Science Foundation of China(Nos.32070940 and 81991491)the China Postdoctoral Science Foundation(No.2021M700115)+2 种基金the Postdoctoral Innovation Talents Support Program(No.BX20220189,China)the Science and Technology Planning Project of Fujian Province(No.2022L3080,China)the CAMS Innovation Fund for Medical Sciences(No.2019RU022,China).
文摘T cell-redirecting bispecific antibodies are specifically designed to bind to tumor-associated antigens,thereby engaging with CD3 on the T cell receptor.This linkage between tumor cells and T cells actively triggers T cell activation and initiates targeted killing of the identified tumor cells.These antibodies have emerged as one of the most promising avenues within tumor immunotherapy.However,despite success in treating hematological malignancies,significant advancements in solid tumors have yet to be explored.In this review,we aim to address the critical challenges associated with T cellredirecting bispecific antibodies and explore novel strategies to overcome these obstacles,with the ultimate goal of expanding the application of this therapy to include solid tumors.
