Advances in antibody engineering have led to the generation of more innovative antibody drugs,such as bispecific antibodies(bs Abs).Following the success associated with blinatumomab,bs Abs have attracted enormous int...Advances in antibody engineering have led to the generation of more innovative antibody drugs,such as bispecific antibodies(bs Abs).Following the success associated with blinatumomab,bs Abs have attracted enormous interest in the field of cancer immunotherapy.By specifically targeting two different antigens,bs Abs reduce the distance between tumor and immune cells,thereby enhancing tumor killing directly.There are several mechanisms of action upon which bs Abs have been exploited.Accumulating experience on checkpoint-based therapy has promoted the clinical transformation of bs Abs targeting immunomodulatory checkpoints.Cadonilimab(PD-1×CTLA-4)is the first approved bs Ab targeting dual inhibitory checkpoints,which confirms the feasibility of bs Abs in immunotherapy.In this review we analyzed the mechanisms by which bs Abs targeting immunomodulatory checkpoints and their emerging applications in cancer immunotherapy.展开更多
Bispecific antibodies (BsAbs) of anti CD3×anti idiotype (Id) to B cell lymphocytic leukemia (CLL) were prepared by chemical conjugation and direct hybridization technique of hybridoma and hybridoma without scr...Bispecific antibodies (BsAbs) of anti CD3×anti idiotype (Id) to B cell lymphocytic leukemia (CLL) were prepared by chemical conjugation and direct hybridization technique of hybridoma and hybridoma without screening markers. The specificity of BsAbs from culture supernatants or ascites was assayed by indirect ELISA and indirect immunoflurescence (IF). The results showed that BsAbs could specifically react with homologous serum IgM from patients with B CLL and cells carrying CD3 marker respectively. Cell combination test and LDH assay demonstrated that BsAb significantly increased the conjugate formation between lymphocyte activated kill (LAK) cells and Daudi cells, and enhanced the cytotoxic activity of LAK cells against Daudi cells.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
Monoclonal antibodies have been successfully incorporated into the current therapeutical landscape of lung cancer in the last decades.Recently,with technological advances,bispecific antibodies(bsAbs)have also shown ro...Monoclonal antibodies have been successfully incorporated into the current therapeutical landscape of lung cancer in the last decades.Recently,with technological advances,bispecific antibodies(bsAbs)have also shown robust efficacy in the treatment of malignant cancers,including lung cancer.These antibodies target two independent epitopes or antigens and have been extensively explored in translational and clinical studies in lung cancer.Here,we outline the mechanisms of action of bsAbs,related clinical data,ongoing clinical trials,and potent novel compounds of various types of bsAbs in clinical studies,especially in lung cancer.We also propose future directions for the clinical development of bsAbs,which might bring a new era of treatment for patients with lung cancer.展开更多
Bispecific antibodies hold significant potential as next-generation biotherapeutics owing to their ability to simultaneously bind to two targets.However,the development of bispecific antibodies as biotherapeutics has ...Bispecific antibodies hold significant potential as next-generation biotherapeutics owing to their ability to simultaneously bind to two targets.However,the development of bispecific antibodies as biotherapeutics has been hindered by the high levels of byproducts produced,including both high molecular weight and low molecular weight variants.In addition,the inevitable expression of homodimers in host cells presents further obstacles to the commercial development of bispecific antibodies as therapeutics.These byproducts,which share similar physicochemical properties with the target,pose several challenges for downstream purification processes.In this study,we present a non-protein A purification platform that employ a one-step polishing chromatography to purify bispecific antibodies.Mixed-mode Capto adhere resin was used to capture the target protein at pH 7.90±0.10,followed by anion exchange chromatography as a polishing step.Overall,the results of this two-step chromatography purification method demonstrated at final product purity of 98%as assessed by size-exclusion high-performance liquid chromatography(SEC-HPLC)and 98%by reversed-phase-high-performance liquid chromatography(RP-HPLC),with residual host cell proteins controlled at 10 ppm and an excellent recovery rate of approximately 60%.This study presents a non-protein A capture platform,offering a simplified,streamlined,and competitive alternative to conventional affinity chromatography.展开更多
In recent years,the development of bispecific antibodies(bsAbs)has been rapid,with many new structures and target combinations being created.The boom in bsAbs has led to the successive issuance of industry guidance fo...In recent years,the development of bispecific antibodies(bsAbs)has been rapid,with many new structures and target combinations being created.The boom in bsAbs has led to the successive issuance of industry guidance for their development in the US and China.However,there is a high degree of similarity in target selection,which could affect the development of diversity in bsAbs.This review presents a classification of various bsAbs for cancer therapy based on structure and target selection and examines the advantages of bsAbs over monoclonal antibodies(mAbs).Through database research,we have identified the preferences of available bsAbs combinations,suggesting rational target selection options and warning of potential wastage of medical resources.We have also compared the US and Chinese guidelines for bsAbs in order to provide a reference for their development.展开更多
Bispecific antibodies are recombinant proteins with novel immunological properties and therapeutic potential. Recombinant protein quality and activity of several bispecific antibodies comprising different variable dom...Bispecific antibodies are recombinant proteins with novel immunological properties and therapeutic potential. Recombinant protein quality and activity of several bispecific antibodies comprising different variable domain combinations with respect to the parental monospecific single chain fragments (scFv) were evaluated after expression in bacteria or mammalian cells. The parental scFv proteins humanized anti-NCAM scFv, murine anti-VEGFR-2 scFv, murine and humanized anti-CD3 scFv, respectively, could successfully be expressed in E. coli, whereas the murine anti-NCAM scFv version could not be reliably detected. Bispecific CD3 × VEGFR-2 and CD3 × NCAM anti-bodies were expressed in the bispecific single chain and the single chain diabody format. However, the diabody derived from the murine anti-NCAM scFv could not efficiently be expressed in E. coli or in mammalian cells. Significant binding of the CD3 × NCAM single chain diabody comprising the humanized version of anti-CD3 and humanized version of anti-NCAM was efficient to both antigens. Nevertheless, binding of the bispecific single chain version to the NCAM antigen was inefficient in comparison to CD3 binding. In conclusion, the data could indicate that the result of scFv expression in bacteria may be predictive for the chances of success for functional expression of more complex bispecific derivatives.展开更多
Bispecific antibodies(bsAbs)hold promises for enhanced therapeutic potential surpassing that of their parental monoclonal antibodies.However,bsAbs pose great challenges in their manufacturing,and one of the common rea...Bispecific antibodies(bsAbs)hold promises for enhanced therapeutic potential surpassing that of their parental monoclonal antibodies.However,bsAbs pose great challenges in their manufacturing,and one of the common reasons is their susceptibility to aggregation.Building on previous studies demonstrating the functionality and potential manufacturability of Fab-scFv format bsAb,this investigation delved into the impact of environmental factors-such as pH,buffer types,ionic strength,protein concentrations,and temperatures-on its stability and the reversal of its self-associated aggregates.Mildly acidic,low-salt conditions were found optimal,ensuring bsAb stability for 30 days even at elevated temperature of 40°C.Furthermore,these conditions facilitated the reversal of its self-associated aggregates to monomers during the initial 7-day incubation period.Our findings underscore the robustness and resilience of Fab-scFv format bsAb,further confirming its potential manufacturability despite its current absence as commercial products.展开更多
The prosperity of the biotherapeutics market reflects the feasibility and effectiveness of therapeutic antibodies for the treatment of cancers,inflammatory disorders,and refractory infections.As drawbacks emerge in cl...The prosperity of the biotherapeutics market reflects the feasibility and effectiveness of therapeutic antibodies for the treatment of cancers,inflammatory disorders,and refractory infections.As drawbacks emerge in clinical trials and practice,such as impeded binding,reduced effector functions,and frequent adverse reactions,modifications of therapeutic antibodies are unprecedently burgeoning in research and development(R&D).These modifications include:①modified glycosylation;②fragment of crystallizable domain(Fc)amino acid alterations;③cross-isotype or cross-subclass exchanges;④antibody–drug conjugates(ADCs);⑤single chain of variable region fragment(scFv)for chimeric antigen receptor T(CAR-T)cells;and⑥bispecific antibodies(bsAbs)in order to promote binding affinity,half-life in circulation,effectiveness toward target cells and,ultimately,to achieve overall improved efficacy.While many achievements have been made around the world in the past decades,China has been playing an active role in this realm,with its great demand for biotherapeutics with R&D potential.This review recapitulates the international progress that has been achieved with modified therapeutic antibodies,and then focuses on that of China in an independent section.展开更多
Objective This study aimed to explore the value of M701,targeting epithelial cell adhesion molecule(EpCAM)and CD3,in the immunotherapy of ovarian cancer ascites by the in vitro assay.Methods The expression of EpCAM in...Objective This study aimed to explore the value of M701,targeting epithelial cell adhesion molecule(EpCAM)and CD3,in the immunotherapy of ovarian cancer ascites by the in vitro assay.Methods The expression of EpCAM in ovarian cancer tissues was analyzed by databases.The EpCAM expression and immune cell infiltration in different foci of ovarian cancer were detected by 8-channel flow cytometry.The toxic effect of M701 on OVCAR3 was tested using the in vitro cytotoxicity assay.The 3D cell culture and drug intervention experiments were performed to evaluate the therapeutic effect of M701 in ovarian cancer specimens.Flow cytometry was used to examine the effect of M701 on the binding of immune cells to tumor cells and the activation capacity of T cells.Results The results of the bioinformatic analysis showed that the expression of EpCAM in ovarian cancer tissue was significantly higher than that in normal ovarian tissue.The 8-channel flow cytometry of clinical samples showed that the EpCAM expression and lymphocyte infiltration were significantly heterogeneous among ovarian cancer patients and lesions at different sites.The in vitro experiment results showed that M701 had a significant killing effect on OVCAR3 cells.M701 also obviously killed primary tumor cells derived from some patients with ovarian cancer ascites.M701 could mediate the binding of CD3^(+)T cells to EpCAM^(+)tumor cells and induce T cell activation in a dose-dependent manner.Conclusion M701 showed significant inhibitory activity on tumor cells derived from ovarian cancer ascites,which had a promising application in immunotherapy for patients with ovarian cancer ascites.展开更多
Selecting an ideal molecular format from diverse structures is a major challenge in developing a bispecific antibody(BsAb).To choose an ideal format of anti-CD3 x anti-transferrin receptor(TfR)bispecific antibodies fo...Selecting an ideal molecular format from diverse structures is a major challenge in developing a bispecific antibody(BsAb).To choose an ideal format of anti-CD3 x anti-transferrin receptor(TfR)bispecific antibodies for clinical application,we constructed TfR bispecific T-cell engager(BiTE)in two extensively applied formats,including single-chain tandem singlechain variable fragments(scFvs)and double-chain diabodies,and evaluated their functional characterizations in vitro.Results demonstrated that TfR-BiTE in both formats directed potent killing of TfR+HepG2 cells.However,compared to two・chain diabodies,scFvs were more efficient in antigen binding and TfR target killing.Furthermore,different domain orders in scFvs would also be evaluated because single-TfR-CD3-His was preferable to single-CD3-TfR-His in immunotherapeutic strategies.Thus,the single-chain tandem TfR-CD3 format was favored for further investigation in cancer therapy.展开更多
Systemic lupus erythematosus(SLE)is characterized by a systemic dysfunction of both the innate and adaptive immune systems,leading to an attack on healthy tissues of the body.During the development of SLE,pathogenic f...Systemic lupus erythematosus(SLE)is characterized by a systemic dysfunction of both the innate and adaptive immune systems,leading to an attack on healthy tissues of the body.During the development of SLE,pathogenic features,such as the formation of autoantibodies against self-nuclear antigens,cause tissue damage including necrosis and fibrosis,with increased expression levels of the typeⅠinterferon-regulated genes.Standard treatments for lupus with immunosuppressants and glucocorticoids are not effective enough but cause side effects.As an alternative,more effective immunotherapies have been developed,including monoclonal and bispecific antibodies that target B cells,T cells,co-stimulatory molecules,cytokines or their receptors,and signaling molecules.Encouraging results have been observed in clinical trials with some of these therapies.Furthermore,a chimeric antigen receptor T cell therapy has emerged as the most effective,safe,and promising treatment option for SLE,as demonstrated by successful pilot studies.Additionally,some emerging evidence suggests that gut microbiota dysbiosis may significantly contribute to the severity of SLE,and the normalization of the gut microbiota through methods such as fecal microbiota transplantation presents new opportunities for effective treatment of SLE.展开更多
BACKGROUND The prognosis for patients with advanced metastatic cervix cancer(MCC)is poor,and this disease continues to pose a considerable therapeutic challenge.Despite the administration of first-line regimens consis...BACKGROUND The prognosis for patients with advanced metastatic cervix cancer(MCC)is poor,and this disease continues to pose a considerable therapeutic challenge.Despite the administration of first-line regimens consisting of cisplatin,paclitaxel,and bevacizumab,survival rates for patients with metastasis remain poor.The emergence of bispecific antibodies(BsAbs)offers a novel treatment option for patients diagnosed with MCC.CASE SUMMARY In this report,we present a patient with MCC who was treated with cadonilimab monotherapy at a dose of 6 mg/kg every two weeks after chemotherapy was proven to be intolerable.The patient exhibited a sustained complete response for a duration of 6 months,demonstrating an optimistic outlook.CONCLUSION This case illustrates the considerable efficacy of cadonilimab for treating advanced MCC.Therefore,BsAb therapy is a promising strategy for effectively treating patients with advanced MCC and should be considered as an option when patients are intolerant to standard chemotherapy.展开更多
The development of immunotherapies for lymphoma has undergone a revolutionary evolution over the past decades. Since the advent of rituximab as the first successful immunotherapy for B-cell non-Hodgkin lymphoma over t...The development of immunotherapies for lymphoma has undergone a revolutionary evolution over the past decades. Since the advent of rituximab as the first successful immunotherapy for B-cell non-Hodgkin lymphoma over two decades ago, a plethora of new immunotherapeutic approaches to treat lymphoma has ensued. Four of the most exciting classes of immunotherapies include:chimeric antigen receptor T-cells, bispecific antibodies, immune checkpoint inhibitors, and vaccines. However, with addition of these novel therapies the appropriate timing of treatment, optimal patient population, duration of therapy, toxicity, and cost must be considered. In this review, we describe the most-promising immunotherapeutic approaches for the treatment of lymphoma in clinical development, specifically focusing on clinical trials performed to date and strategies for improvement.展开更多
Treatment strategies for inflammatory bowel disease(IBD)are rapidly evolving with the development of biologics and small molecule drugs(SMDs).However,these drugs are not guaranteed to be effective in all patients,and ...Treatment strategies for inflammatory bowel disease(IBD)are rapidly evolving with the development of biologics and small molecule drugs(SMDs).However,these drugs are not guaranteed to be effective in all patients,and a“ceiling effect”of biologic monotherapy may occur.This issue highlights an unmet need for optimizing the use of biologics and predicting therapeutic responses.Thus,the development of new drugs with novel mechanisms of action is urgently needed for patients with primary nonresponse and secondary loss of response to conventional biologics and SMDs.In addition,combining different biologics or SMDs has been proposed as a novel strategy to enhance treatment efficacy in IBD,which theoretically has multidimensional anti-inflammatory potential.Based on the current evidence available for IBD,dual targeted therapy may be a promising strategy for refractory IBD patients who have failed in multiple biologic treatments or who have extraintestinal manifestation.Additionally,identifying the subgroup of IBD patients who are responding to biological combination therapies is also equally important in stable disease remission.In this review,we summarize the newly developed biologics and SMDs and the current status of biologics/SMDs to highlight the development of individualized treatment in IBD.展开更多
Follicular lymphoma(FL)is the most common low-grade lymphoma,and although nodal FL is highly responsive to treatment,the majority of patients relapse repeatedly,and the disease has been incurable with a poor prognosis...Follicular lymphoma(FL)is the most common low-grade lymphoma,and although nodal FL is highly responsive to treatment,the majority of patients relapse repeatedly,and the disease has been incurable with a poor prognosis.However,primary FL of the gastrointestinal tract has been increasingly detected in Japan,especially due to recent advances in small bowel endoscopy and increased opportunities for endoscopic examinations and endoscopic diagnosis.However,many cases are detected at an early stage,and the prognosis is good in many cases.In contrast,in Europe and the United States,gastrointestinal FL has long been considered to be present in 12%-24%of Stage-IV patients,and the number of advanced gastrointestinal cases is expected to increase.This editorial provides an overview of the recent therapeutic advances in nodal FL,including antibody-targeted therapy,bispecific antibody therapy,epigenetic modulation,and chimeric antigen receptor T-cell therapy,and reviews the latest therapeutic manuscripts published in the past year.Based on an understanding of the therapeutic advances in nodal FL,we also discuss future possibilities for gastroenterologists to treat gastrointestinal FL,especially in advanced cases.展开更多
The CD19-targeting bispecific T-cell engager blinatumomab has shown remarkable efficacy in patients with relapsed/refractory B-cell precursor acute lymphoblastic leukemia.However,several studies showed that blinatumom...The CD19-targeting bispecific T-cell engager blinatumomab has shown remarkable efficacy in patients with relapsed/refractory B-cell precursor acute lymphoblastic leukemia.However,several studies showed that blinatumomab has a short plasma half-life due to its low molecular weight,and thus its clinical use is limited.Furthermore,multiple trials have shown that approximately 30%of blinatumomab-relapsed cases are characterized by CD19 negative leukemic cells.Here,we design and characterize two novel antibodies,A-319 and A-2019.Blinatumomab and A-319 are CD3/CD19 bispecific antibodies with different molecular sizes and structures,and A-2019 is a novel CD3/CD19/CD20 trispecific antibody with an additional anti-CD20 function.Our in vitro,ex vivo,and in vivo experiments demonstrated that A-319 and A-2019 are potent antitumor agents and capable of recruiting CD3 positive T cells,enhancing T-cell function,mediating B-cell depletion,and eventually inhibiting tumor growth in Raji xenograft models.The two molecules are complementary in terms of efficacy and specificity profile.The activity of A-319 demonstrated superior to that of A-2019,whereas A-2019 has an additional capability to target CD20 in cells missing CD19,suggesting its potential function against CD19 weak or negative CD20 positive leukemic cells.展开更多
Bispecific antibody‒drug conjugates(BsADCs)represent an innovative therapeutic category amalgamating the merits of antibody‒drug conjugates(ADCs)and bispecific antibodies(BsAbs).Positioned as the next-generation ADC a...Bispecific antibody‒drug conjugates(BsADCs)represent an innovative therapeutic category amalgamating the merits of antibody‒drug conjugates(ADCs)and bispecific antibodies(BsAbs).Positioned as the next-generation ADC approach,BsADCs hold promise for ameliorating extant clinical challenges associated with ADCs,particularly pertaining to issues such as poor internalization,off-target toxicity,and drug resistance.Presently,ten BsADCs are undergoing clinical trials,and initial findings underscore the imperative for ongoing refinement.This review initially delves into specific design considerations for BsADCs,encompassing target selection,antibody formats,and the linker–payload complex.Subsequent sections delineate the extant progress and challenges encountered by BsADCs,illustrated through pertinent case studies.The amalgamation of BsAbs with ADCs offers a prospective solution to prevailing clinical limitations of ADCs.Nevertheless,the symbiotic interplay among BsAb,linker,and payload necessitates further optimizations and coordination beyond a simplistic“1+1”to effectively surmount the extant challenges facing the BsADC domain.展开更多
Antibody–drug conjugates(ADCs)are biologically targeted drugs composed of antibodies and cytotoxic drugs connected by linkers.These innovative compounds enable precise drug delivery to tumor cells,minimizing harm to ...Antibody–drug conjugates(ADCs)are biologically targeted drugs composed of antibodies and cytotoxic drugs connected by linkers.These innovative compounds enable precise drug delivery to tumor cells,minimizing harm to normal tissues and offering excellent prospects for cancer treatment.However,monoclonal antibody-based ADCs still present challenges,especially in terms of balancing efficacy and safety.Bispecific antibodies are alternatives to monoclonal antibodies and exhibit superior internalization and selectivity,producing ADCs with increased safety and therapeutic efficacy.In this review,we present available evidence and future prospects regarding the use of bispecific ADCs for cancer treatment,including a comprehensive overview of bispecific ADCs that are currently in clinical trials.We offer insights into the future development of bispecific ADCs to provide novel strategies for cancer treatment.展开更多
The anti-amyloid-j(anti-Aβ)fibrils and soluble oligomers antibody aducanumab were approved to effectively slow down the progression of A lzheimer's disease(AD)at higher doses in 2019,reaffirming the therapeutic e...The anti-amyloid-j(anti-Aβ)fibrils and soluble oligomers antibody aducanumab were approved to effectively slow down the progression of A lzheimer's disease(AD)at higher doses in 2019,reaffirming the therapeutic effects of targeting the core pathology of AD.A timely and accurate diagnosis in the prodromal or pre-dementia stage of AD is essential for patient recruitment,stratifcation,and monitoring of treat ment effects.A D core biomarkers amyloid-B(Aβ1-42),total tau(t-tau),and phosphorylated tau(p-tau)have been clinically validated to reflect AD-type pathological changes through cerebrospinal fuid(CSF)measurement or positron-emission to-mography(PET)and found to have high diagnostic performance for AD identification in the stage of mild cognitive impairment.The development of ultrasensitive immunoassay technology enables AD pathological proteins such as tau and neurofilament light(NFL)to be measured in blood samples.However,combined biomarker detection or targeting multiple biomarkers in immunoassays will increase detection sensitivity and specifcity and improve diagnostic accuracy..This review summarizes and analyzes the performance of current detection methods for early diagnosis of AD,and provides a concept of detection method based on multiple biomarkers instead of a single target,which may become a potential tool for early diagnosis of AD in the future.展开更多
基金supported by the National Natural Science Foundation of China(Grant Nos.82130077 and 81961128025)。
文摘Advances in antibody engineering have led to the generation of more innovative antibody drugs,such as bispecific antibodies(bs Abs).Following the success associated with blinatumomab,bs Abs have attracted enormous interest in the field of cancer immunotherapy.By specifically targeting two different antigens,bs Abs reduce the distance between tumor and immune cells,thereby enhancing tumor killing directly.There are several mechanisms of action upon which bs Abs have been exploited.Accumulating experience on checkpoint-based therapy has promoted the clinical transformation of bs Abs targeting immunomodulatory checkpoints.Cadonilimab(PD-1×CTLA-4)is the first approved bs Ab targeting dual inhibitory checkpoints,which confirms the feasibility of bs Abs in immunotherapy.In this review we analyzed the mechanisms by which bs Abs targeting immunomodulatory checkpoints and their emerging applications in cancer immunotherapy.
文摘Bispecific antibodies (BsAbs) of anti CD3×anti idiotype (Id) to B cell lymphocytic leukemia (CLL) were prepared by chemical conjugation and direct hybridization technique of hybridoma and hybridoma without screening markers. The specificity of BsAbs from culture supernatants or ascites was assayed by indirect ELISA and indirect immunoflurescence (IF). The results showed that BsAbs could specifically react with homologous serum IgM from patients with B CLL and cells carrying CD3 marker respectively. Cell combination test and LDH assay demonstrated that BsAb significantly increased the conjugate formation between lymphocyte activated kill (LAK) cells and Daudi cells, and enhanced the cytotoxic activity of LAK cells against Daudi cells.
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.
基金sponsored by grants from the National Natural Science Foundation of China(Nos.82172869 and 81972167)the Program of Shanghai Academic Research Leader(No.21XD1423200)the program of Shanghai Shenkang Hospital Development Center(No.SHDC12019133).
文摘Monoclonal antibodies have been successfully incorporated into the current therapeutical landscape of lung cancer in the last decades.Recently,with technological advances,bispecific antibodies(bsAbs)have also shown robust efficacy in the treatment of malignant cancers,including lung cancer.These antibodies target two independent epitopes or antigens and have been extensively explored in translational and clinical studies in lung cancer.Here,we outline the mechanisms of action of bsAbs,related clinical data,ongoing clinical trials,and potent novel compounds of various types of bsAbs in clinical studies,especially in lung cancer.We also propose future directions for the clinical development of bsAbs,which might bring a new era of treatment for patients with lung cancer.
文摘Bispecific antibodies hold significant potential as next-generation biotherapeutics owing to their ability to simultaneously bind to two targets.However,the development of bispecific antibodies as biotherapeutics has been hindered by the high levels of byproducts produced,including both high molecular weight and low molecular weight variants.In addition,the inevitable expression of homodimers in host cells presents further obstacles to the commercial development of bispecific antibodies as therapeutics.These byproducts,which share similar physicochemical properties with the target,pose several challenges for downstream purification processes.In this study,we present a non-protein A purification platform that employ a one-step polishing chromatography to purify bispecific antibodies.Mixed-mode Capto adhere resin was used to capture the target protein at pH 7.90±0.10,followed by anion exchange chromatography as a polishing step.Overall,the results of this two-step chromatography purification method demonstrated at final product purity of 98%as assessed by size-exclusion high-performance liquid chromatography(SEC-HPLC)and 98%by reversed-phase-high-performance liquid chromatography(RP-HPLC),with residual host cell proteins controlled at 10 ppm and an excellent recovery rate of approximately 60%.This study presents a non-protein A capture platform,offering a simplified,streamlined,and competitive alternative to conventional affinity chromatography.
基金supported by the National Key Research and Development Program of China(2022YFA1303803)National Natural Science Foundation of China(82073701)+1 种基金the Project Program of State Key Laboratory of Natural Medicines,China Pharmaceutical University(SKLNMZZ202209)supported by“Double First-Class”University Project(CPU2022PZQ07,China)。
文摘In recent years,the development of bispecific antibodies(bsAbs)has been rapid,with many new structures and target combinations being created.The boom in bsAbs has led to the successive issuance of industry guidance for their development in the US and China.However,there is a high degree of similarity in target selection,which could affect the development of diversity in bsAbs.This review presents a classification of various bsAbs for cancer therapy based on structure and target selection and examines the advantages of bsAbs over monoclonal antibodies(mAbs).Through database research,we have identified the preferences of available bsAbs combinations,suggesting rational target selection options and warning of potential wastage of medical resources.We have also compared the US and Chinese guidelines for bsAbs in order to provide a reference for their development.
基金financial support of AK by a grant of the Clotten-Stiftung,Freiburg,GermanyPPM was supported by the Deutsche Forschungsgemeinschaft DFG grant SFB599.
文摘Bispecific antibodies are recombinant proteins with novel immunological properties and therapeutic potential. Recombinant protein quality and activity of several bispecific antibodies comprising different variable domain combinations with respect to the parental monospecific single chain fragments (scFv) were evaluated after expression in bacteria or mammalian cells. The parental scFv proteins humanized anti-NCAM scFv, murine anti-VEGFR-2 scFv, murine and humanized anti-CD3 scFv, respectively, could successfully be expressed in E. coli, whereas the murine anti-NCAM scFv version could not be reliably detected. Bispecific CD3 × VEGFR-2 and CD3 × NCAM anti-bodies were expressed in the bispecific single chain and the single chain diabody format. However, the diabody derived from the murine anti-NCAM scFv could not efficiently be expressed in E. coli or in mammalian cells. Significant binding of the CD3 × NCAM single chain diabody comprising the humanized version of anti-CD3 and humanized version of anti-NCAM was efficient to both antigens. Nevertheless, binding of the bispecific single chain version to the NCAM antigen was inefficient in comparison to CD3 binding. In conclusion, the data could indicate that the result of scFv expression in bacteria may be predictive for the chances of success for functional expression of more complex bispecific derivatives.
基金supported by Agency for Science,Technology,and Research(A*STAR)BMRC Central Research Fund.
文摘Bispecific antibodies(bsAbs)hold promises for enhanced therapeutic potential surpassing that of their parental monoclonal antibodies.However,bsAbs pose great challenges in their manufacturing,and one of the common reasons is their susceptibility to aggregation.Building on previous studies demonstrating the functionality and potential manufacturability of Fab-scFv format bsAb,this investigation delved into the impact of environmental factors-such as pH,buffer types,ionic strength,protein concentrations,and temperatures-on its stability and the reversal of its self-associated aggregates.Mildly acidic,low-salt conditions were found optimal,ensuring bsAb stability for 30 days even at elevated temperature of 40°C.Furthermore,these conditions facilitated the reversal of its self-associated aggregates to monomers during the initial 7-day incubation period.Our findings underscore the robustness and resilience of Fab-scFv format bsAb,further confirming its potential manufacturability despite its current absence as commercial products.
基金This study was supported by the National Basic Research Program of China(2015CB553701)the National High Technology Research and Development Program of China(2019ZX09732001).
文摘The prosperity of the biotherapeutics market reflects the feasibility and effectiveness of therapeutic antibodies for the treatment of cancers,inflammatory disorders,and refractory infections.As drawbacks emerge in clinical trials and practice,such as impeded binding,reduced effector functions,and frequent adverse reactions,modifications of therapeutic antibodies are unprecedently burgeoning in research and development(R&D).These modifications include:①modified glycosylation;②fragment of crystallizable domain(Fc)amino acid alterations;③cross-isotype or cross-subclass exchanges;④antibody–drug conjugates(ADCs);⑤single chain of variable region fragment(scFv)for chimeric antigen receptor T(CAR-T)cells;and⑥bispecific antibodies(bsAbs)in order to promote binding affinity,half-life in circulation,effectiveness toward target cells and,ultimately,to achieve overall improved efficacy.While many achievements have been made around the world in the past decades,China has been playing an active role in this realm,with its great demand for biotherapeutics with R&D potential.This review recapitulates the international progress that has been achieved with modified therapeutic antibodies,and then focuses on that of China in an independent section.
基金This work was supported by the National Key Research&Development Program of China(No.2021YFC2701402).
文摘Objective This study aimed to explore the value of M701,targeting epithelial cell adhesion molecule(EpCAM)and CD3,in the immunotherapy of ovarian cancer ascites by the in vitro assay.Methods The expression of EpCAM in ovarian cancer tissues was analyzed by databases.The EpCAM expression and immune cell infiltration in different foci of ovarian cancer were detected by 8-channel flow cytometry.The toxic effect of M701 on OVCAR3 was tested using the in vitro cytotoxicity assay.The 3D cell culture and drug intervention experiments were performed to evaluate the therapeutic effect of M701 in ovarian cancer specimens.Flow cytometry was used to examine the effect of M701 on the binding of immune cells to tumor cells and the activation capacity of T cells.Results The results of the bioinformatic analysis showed that the expression of EpCAM in ovarian cancer tissue was significantly higher than that in normal ovarian tissue.The 8-channel flow cytometry of clinical samples showed that the EpCAM expression and lymphocyte infiltration were significantly heterogeneous among ovarian cancer patients and lesions at different sites.The in vitro experiment results showed that M701 had a significant killing effect on OVCAR3 cells.M701 also obviously killed primary tumor cells derived from some patients with ovarian cancer ascites.M701 could mediate the binding of CD3^(+)T cells to EpCAM^(+)tumor cells and induce T cell activation in a dose-dependent manner.Conclusion M701 showed significant inhibitory activity on tumor cells derived from ovarian cancer ascites,which had a promising application in immunotherapy for patients with ovarian cancer ascites.
基金National Natural Science Foundation of China(No.31570937 and No.81871391)Natural Science Foundation of Hubei Province of China(No.2017CFB707)+1 种基金the Fundamental Research Funds for the Central Universities of China(No.HUST:2018KFYYXJJ086)Graduates'Innovation Foundation of Huazhong University of Science and Technology(No.5003510001).
文摘Selecting an ideal molecular format from diverse structures is a major challenge in developing a bispecific antibody(BsAb).To choose an ideal format of anti-CD3 x anti-transferrin receptor(TfR)bispecific antibodies for clinical application,we constructed TfR bispecific T-cell engager(BiTE)in two extensively applied formats,including single-chain tandem singlechain variable fragments(scFvs)and double-chain diabodies,and evaluated their functional characterizations in vitro.Results demonstrated that TfR-BiTE in both formats directed potent killing of TfR+HepG2 cells.However,compared to two・chain diabodies,scFvs were more efficient in antigen binding and TfR target killing.Furthermore,different domain orders in scFvs would also be evaluated because single-TfR-CD3-His was preferable to single-CD3-TfR-His in immunotherapeutic strategies.Thus,the single-chain tandem TfR-CD3 format was favored for further investigation in cancer therapy.
基金funded by the Russian Science Foundation Grant No.21-74-10154 to A.K.
文摘Systemic lupus erythematosus(SLE)is characterized by a systemic dysfunction of both the innate and adaptive immune systems,leading to an attack on healthy tissues of the body.During the development of SLE,pathogenic features,such as the formation of autoantibodies against self-nuclear antigens,cause tissue damage including necrosis and fibrosis,with increased expression levels of the typeⅠinterferon-regulated genes.Standard treatments for lupus with immunosuppressants and glucocorticoids are not effective enough but cause side effects.As an alternative,more effective immunotherapies have been developed,including monoclonal and bispecific antibodies that target B cells,T cells,co-stimulatory molecules,cytokines or their receptors,and signaling molecules.Encouraging results have been observed in clinical trials with some of these therapies.Furthermore,a chimeric antigen receptor T cell therapy has emerged as the most effective,safe,and promising treatment option for SLE,as demonstrated by successful pilot studies.Additionally,some emerging evidence suggests that gut microbiota dysbiosis may significantly contribute to the severity of SLE,and the normalization of the gut microbiota through methods such as fecal microbiota transplantation presents new opportunities for effective treatment of SLE.
文摘BACKGROUND The prognosis for patients with advanced metastatic cervix cancer(MCC)is poor,and this disease continues to pose a considerable therapeutic challenge.Despite the administration of first-line regimens consisting of cisplatin,paclitaxel,and bevacizumab,survival rates for patients with metastasis remain poor.The emergence of bispecific antibodies(BsAbs)offers a novel treatment option for patients diagnosed with MCC.CASE SUMMARY In this report,we present a patient with MCC who was treated with cadonilimab monotherapy at a dose of 6 mg/kg every two weeks after chemotherapy was proven to be intolerable.The patient exhibited a sustained complete response for a duration of 6 months,demonstrating an optimistic outlook.CONCLUSION This case illustrates the considerable efficacy of cadonilimab for treating advanced MCC.Therefore,BsAb therapy is a promising strategy for effectively treating patients with advanced MCC and should be considered as an option when patients are intolerant to standard chemotherapy.
基金supported by NIH grants(Grant No.R01CA136934,and R01 CA193167)
文摘The development of immunotherapies for lymphoma has undergone a revolutionary evolution over the past decades. Since the advent of rituximab as the first successful immunotherapy for B-cell non-Hodgkin lymphoma over two decades ago, a plethora of new immunotherapeutic approaches to treat lymphoma has ensued. Four of the most exciting classes of immunotherapies include:chimeric antigen receptor T-cells, bispecific antibodies, immune checkpoint inhibitors, and vaccines. However, with addition of these novel therapies the appropriate timing of treatment, optimal patient population, duration of therapy, toxicity, and cost must be considered. In this review, we describe the most-promising immunotherapeutic approaches for the treatment of lymphoma in clinical development, specifically focusing on clinical trials performed to date and strategies for improvement.
基金Jiangsu Provincial Health Commission,No.M2021013the Science Foundation of Jinling Hospital,No.YYMS2021035.
文摘Treatment strategies for inflammatory bowel disease(IBD)are rapidly evolving with the development of biologics and small molecule drugs(SMDs).However,these drugs are not guaranteed to be effective in all patients,and a“ceiling effect”of biologic monotherapy may occur.This issue highlights an unmet need for optimizing the use of biologics and predicting therapeutic responses.Thus,the development of new drugs with novel mechanisms of action is urgently needed for patients with primary nonresponse and secondary loss of response to conventional biologics and SMDs.In addition,combining different biologics or SMDs has been proposed as a novel strategy to enhance treatment efficacy in IBD,which theoretically has multidimensional anti-inflammatory potential.Based on the current evidence available for IBD,dual targeted therapy may be a promising strategy for refractory IBD patients who have failed in multiple biologic treatments or who have extraintestinal manifestation.Additionally,identifying the subgroup of IBD patients who are responding to biological combination therapies is also equally important in stable disease remission.In this review,we summarize the newly developed biologics and SMDs and the current status of biologics/SMDs to highlight the development of individualized treatment in IBD.
文摘Follicular lymphoma(FL)is the most common low-grade lymphoma,and although nodal FL is highly responsive to treatment,the majority of patients relapse repeatedly,and the disease has been incurable with a poor prognosis.However,primary FL of the gastrointestinal tract has been increasingly detected in Japan,especially due to recent advances in small bowel endoscopy and increased opportunities for endoscopic examinations and endoscopic diagnosis.However,many cases are detected at an early stage,and the prognosis is good in many cases.In contrast,in Europe and the United States,gastrointestinal FL has long been considered to be present in 12%-24%of Stage-IV patients,and the number of advanced gastrointestinal cases is expected to increase.This editorial provides an overview of the recent therapeutic advances in nodal FL,including antibody-targeted therapy,bispecific antibody therapy,epigenetic modulation,and chimeric antigen receptor T-cell therapy,and reviews the latest therapeutic manuscripts published in the past year.Based on an understanding of the therapeutic advances in nodal FL,we also discuss future possibilities for gastroenterologists to treat gastrointestinal FL,especially in advanced cases.
基金funded by the National Natural Science Foundation of China(Nos.81670147,81570178,Antrag M-0377)Shanghai Municipal Education Commission-Major Project for Scientific Research and Innovation Plan of Natural Science(No.2021-01-07-00-02-E00091)Gaofeng Clinical Medicine Grant Support of Shanghai Municipal Education(No.20172002).
文摘The CD19-targeting bispecific T-cell engager blinatumomab has shown remarkable efficacy in patients with relapsed/refractory B-cell precursor acute lymphoblastic leukemia.However,several studies showed that blinatumomab has a short plasma half-life due to its low molecular weight,and thus its clinical use is limited.Furthermore,multiple trials have shown that approximately 30%of blinatumomab-relapsed cases are characterized by CD19 negative leukemic cells.Here,we design and characterize two novel antibodies,A-319 and A-2019.Blinatumomab and A-319 are CD3/CD19 bispecific antibodies with different molecular sizes and structures,and A-2019 is a novel CD3/CD19/CD20 trispecific antibody with an additional anti-CD20 function.Our in vitro,ex vivo,and in vivo experiments demonstrated that A-319 and A-2019 are potent antitumor agents and capable of recruiting CD3 positive T cells,enhancing T-cell function,mediating B-cell depletion,and eventually inhibiting tumor growth in Raji xenograft models.The two molecules are complementary in terms of efficacy and specificity profile.The activity of A-319 demonstrated superior to that of A-2019,whereas A-2019 has an additional capability to target CD20 in cells missing CD19,suggesting its potential function against CD19 weak or negative CD20 positive leukemic cells.
基金This review was supported by the National Natural Science Foundation of China(82073318)Sichuan Science and Technology Program(2019YFS0003,China)the Support Program of Science&Technology Department of Sichuan Provincial(2023YFSY0046 and 2022NSFSC1365,China).
文摘Bispecific antibody‒drug conjugates(BsADCs)represent an innovative therapeutic category amalgamating the merits of antibody‒drug conjugates(ADCs)and bispecific antibodies(BsAbs).Positioned as the next-generation ADC approach,BsADCs hold promise for ameliorating extant clinical challenges associated with ADCs,particularly pertaining to issues such as poor internalization,off-target toxicity,and drug resistance.Presently,ten BsADCs are undergoing clinical trials,and initial findings underscore the imperative for ongoing refinement.This review initially delves into specific design considerations for BsADCs,encompassing target selection,antibody formats,and the linker–payload complex.Subsequent sections delineate the extant progress and challenges encountered by BsADCs,illustrated through pertinent case studies.The amalgamation of BsAbs with ADCs offers a prospective solution to prevailing clinical limitations of ADCs.Nevertheless,the symbiotic interplay among BsAb,linker,and payload necessitates further optimizations and coordination beyond a simplistic“1+1”to effectively surmount the extant challenges facing the BsADC domain.
基金supported by the National Natural Science Foundation of China(Nos.32070940 and 81991491)China Postdoctoral Science Foundation(No.2021M700115)+2 种基金Postdoctoral Innovation Talents Support Program(No.BX20220189)CAMS Innovation Fund for Medical Sciences(No.2019RU022)Fundamental Research Funds for the Central Universities(No.20720220006).
文摘Antibody–drug conjugates(ADCs)are biologically targeted drugs composed of antibodies and cytotoxic drugs connected by linkers.These innovative compounds enable precise drug delivery to tumor cells,minimizing harm to normal tissues and offering excellent prospects for cancer treatment.However,monoclonal antibody-based ADCs still present challenges,especially in terms of balancing efficacy and safety.Bispecific antibodies are alternatives to monoclonal antibodies and exhibit superior internalization and selectivity,producing ADCs with increased safety and therapeutic efficacy.In this review,we present available evidence and future prospects regarding the use of bispecific ADCs for cancer treatment,including a comprehensive overview of bispecific ADCs that are currently in clinical trials.We offer insights into the future development of bispecific ADCs to provide novel strategies for cancer treatment.
基金the National Natural Science Founda-tion of China(Grant No.81971025)the Startup Fund of Huazhong University of Science and Technology(Grant No.0214187096).
文摘The anti-amyloid-j(anti-Aβ)fibrils and soluble oligomers antibody aducanumab were approved to effectively slow down the progression of A lzheimer's disease(AD)at higher doses in 2019,reaffirming the therapeutic effects of targeting the core pathology of AD.A timely and accurate diagnosis in the prodromal or pre-dementia stage of AD is essential for patient recruitment,stratifcation,and monitoring of treat ment effects.A D core biomarkers amyloid-B(Aβ1-42),total tau(t-tau),and phosphorylated tau(p-tau)have been clinically validated to reflect AD-type pathological changes through cerebrospinal fuid(CSF)measurement or positron-emission to-mography(PET)and found to have high diagnostic performance for AD identification in the stage of mild cognitive impairment.The development of ultrasensitive immunoassay technology enables AD pathological proteins such as tau and neurofilament light(NFL)to be measured in blood samples.However,combined biomarker detection or targeting multiple biomarkers in immunoassays will increase detection sensitivity and specifcity and improve diagnostic accuracy..This review summarizes and analyzes the performance of current detection methods for early diagnosis of AD,and provides a concept of detection method based on multiple biomarkers instead of a single target,which may become a potential tool for early diagnosis of AD in the future.
