Antibody Therapeutics最新文献

{"title":"Application of a novel LC-MS-based proteomic workflow for host cell protein monitoring and risk control in biotherapeutics.","authors":"Xiaojuan Yu, Jianhui Cheng, Qianchuan Lv, Feiyu Wang, Jianzhong Hu, Xinxin Li, Chuanfei Yu","doi":"10.1093/abt/tbag013","DOIUrl":"10.1093/abt/tbag013","url":null,"abstract":"<p><strong>Background: </strong>Comprehensive monitoring of host cell proteins (HCPs) is essential for ensuring the quality, safety, and consistency of antibody therapeutics. Conventional enzyme-linked immunosorbent assay-based assays provide only total HCP burden and lack the resolution to identify individual, functionally active impurities that may affect product stability or efficacy. To address this limitation, we developed a versatile, high-sensitivity mass spectrometry-based workflow capable of molecular-level HCP characterization and risk assessment.</p><p><strong>Methods: </strong>We developed and evaluated a sub-ppm, easy-to-use liquid chromatography-mass spectrometry-based proteomics workflow for the identification and relative quantitation of HCPs across downstream purification and final formulation, which offers molecular-specific, risk-based process characterization and regulatory-aligned quality assurance for antibody therapeutics.</p><p><strong>Results: </strong>The workflow demonstrated sub-ppm sensitivity and excellent reproducibility, enabling comprehensive tracking of HCP clearance across the purification process. Scale-up from 100 to 500 L led to higher total HCP levels and moderately increased species diversity, indicating reduced clearance efficiency and altered impurity profiles. Mechanistic interpretation linked transient thioredoxin and thioredoxin reductase presence to disulfide bond reduction and nonreducing capillary electrophoresis-sodium dodecyl sulfate purity fluctuations. In the formulation stage, lipoprotein lipase was identified as a key contributor to polysorbate 80 (PS80) degradation, consistent with accelerated stability results, leading to successful process optimization.</p><p><strong>Conclusions: </strong>This study establishes a comprehensive, sub-ppm data-independent acquisition (DIA)-mass spectrometry (MS) platform for HCP monitoring in antibody therapeutics, bridging analytical characterization with process and formulation control. The workflow enables molecular-level understanding of impurity behavior, supports proactive risk mitigation, and enhances overall process consistency, representing a significant advancement in HCP management and biotherapeutic quality assurance.</p>","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":"9 2","pages":"159-167"},"PeriodicalIF":4.8,"publicationDate":"2026-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13175985/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147964834","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Affinity-optimized TROP2 antibodies support potent antitumor activity in antibody-drug conjugates.","authors":"Aiko Yamaguchi, Junping Hong, Leike Li, Kiyotaka Kobayashi, Chisato M Yamazaki, Summer Y Y Ha, Yasuaki Anami, Wei Xiong, Junquan Liu, Zhiqiang An, Ningyan Zhang, Kyoji Tsuchikama","doi":"10.1093/abt/tbag011","DOIUrl":"10.1093/abt/tbag011","url":null,"abstract":"<p><strong>Background: </strong>Trophoblast cell surface antigen 2 (TROP2) is frequently overexpressed in epithelial tumors and is associated with poor prognosis, making it an attractive therapeutic target. Antibody-drug conjugates (ADCs) directed against TROP2 show clinical benefit, but expression in normal tissues such as skin raises concerns about on-target, off-tumor toxicity. Strategies that improve antitumor efficacy without increasing toxicity are needed.</p><p><strong>Methods: </strong>Using an in-house phage library, we identified and characterized a novel fully human monoclonal antibody recognizing a unique conformational epitope of TROP2 with reduced binding affinity. This antibody was engineered into homogeneous ADCs carrying auristatin and/or duocarmycin payloads. Comparative studies with a surrogate of sacituzumab govitecan were performed in TROP2-expressing tumor models.</p><p><strong>Results: </strong>The novel ADCs demonstrated remarkable antitumor activity in mouse xenograft and syngeneic tumor models. Despite lower binding affinity, the novel antibody exhibited potent efficacy, suggesting that epitope selection and affinity tuning can be leveraged to enhance therapeutic outcomes.</p><p><strong>Conclusions: </strong>Novel anti-TROP2 ADCs offer a promising approach for assuring efficacy while potentially mitigating toxicity. Optimization of antibody binding properties may enable the development of safer and more effective TROP2-targeted therapeutics.</p>","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":"9 2","pages":"127-138"},"PeriodicalIF":4.8,"publicationDate":"2026-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13070557/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147677355","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Novel ADC and γδ T cell engager targeting CDH17 for the therapy of gastrointestinal cancers.","authors":"Mingcan Yu, Guangmao Mu, Peng Chen, Honglei Bi, Fulai Zhou, Zhengxia Zha, Sheng Huang, Hao Jiang, Ying Jin, Yuanting Chen, Mark L Chiu, Di Zhang","doi":"10.1093/abt/tbag012","DOIUrl":"10.1093/abt/tbag012","url":null,"abstract":"<p><strong>Background: </strong>Cadherin-17 (CDH17) is a cell-adhesion molecule physiologically expressed along the intestinal epithelial tight junctions. Aberrant overexpression of CDH17 in gastrointestinal (GI) cancers promotes tumor growth and metastasis and is associated with poor patient prognosis. Due to its restricted expression in normal tissues and strong association with malignancy, CDH17 represents an emerging therapeutic target for GI tract cancers.</p><p><strong>Methods: </strong>A high-affinity anti-CDH17 monoclonal antibody (TAVO307) was generated and conjugated with auristatin-derived cytotoxic payloads to obtain CDH17-directed antibody-drug conjugates (ADCs). In parallel, a VHH antibody capable of activating γδ T cell receptors from both Vδ1 and Vδ2 subsets was identified and combined with the anti-CDH17 antibody to generate CDH17-targeted T cell engager (TCE) to recruit γδ T cells, which are abundant in the intestinal mucosa and play a critical role in tumor immunosurveillance. An attenuated interleukin-15 (IL-15) fused with the IL-15 receptor α sushi domain was further incorporated on TCE to enhance γδ T cell expansion and activation.</p><p><strong>Results: </strong>CDH17-based ADCs exhibited potent and selective cytotoxicity in multiple GI cancer cell lines and significant tumor regression in xenograft models. The CDH17 γδ TCE induced tumor antigen-dependent γδ T cell degranulation and redirected both Vδ1 and Vδ2 T cells to effectively kill CDH17-expressing cancer cells. IL-15 fusion further augmented γδ T cell expansion and activation.</p><p><strong>Conclusions: </strong>Both CDH17-targeted ADCs and γδ TCEs demonstrated promising potency and efficacy to control GI cancers. They could offer complementary therapeutic options that could be used in combination therapy.</p>","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":"9 3","pages":"230-247"},"PeriodicalIF":4.8,"publicationDate":"2026-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13317742/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148362731","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Systematic functional screening of immunoreceptor tyrosine-based inhibitory motif domains identifies potent inhibitory modules for chimeric antigen receptor-T.","authors":"Ting Yang, Min Gao, Yufang Sun, Zhiwei Li, Zijun Xie, Ming Zou, Mi Deng","doi":"10.1093/abt/tbag010","DOIUrl":"10.1093/abt/tbag010","url":null,"abstract":"<p><strong>Background: </strong>Chimeric antigen receptor (CAR) T-cell therapy has revolutionized the treatment of hematologic malignancies but remains limited by on-target, off-tumor toxicity and the lack of precise mechanisms to restrain aberrant activation. Inhibitory CARs (iCARs) offer a logic-gated strategy to suppress unwanted immune responses, yet optimization of inhibitory signaling modules has been hindered by an incomplete understanding of immunoreceptor tyrosine-based inhibitory motif (ITIM) diversity.</p><p><strong>Methods: </strong>We established a systematic screening platform to evaluate ITIM-containing intracellular domains (ICDs) from 35 candidate receptors. Using nuclear factor of activated T cells (NFAT) reporter assays across multiple activation contexts, we quantified the ability of each domain to counteract iimmunoreceptor tyrosine-based activating motif (ITAM)-mediated signaling. Intracellular truncated analyses tested the requirement of intact cytoplasmic motifs, and the most potent ITIMs were incorporated into iCAR architectures to assess their capacity to inhibit CAR-driven effector responses.</p><p><strong>Results: </strong>We identified 12 ITIM domains that strongly suppressed NFAT signaling across diverse activation states. Their inhibitory activity required intact ITIM motifs. When integrated into iCAR constructs, these domains conferred markedly greater suppression of CAR-induced activation compared with conventional programmed cell death protein 1 (PD-1)-based designs. Among them, the SIGLEC9-derived ITIM exhibited the most potent inhibition, significantly reducing CAR-T effector function and protecting target cells from cytotoxicity <i>in vitro</i>.</p><p><strong>Conclusions: </strong>We identified distinct intracellular modules that effectively suppress CAR-mediated activation, with the SIGLEC9-derived ITIM exhibiting the strongest inhibition. Leveraging domain-specific inhibitory strength enables precise and modular control of T-cell activation, thereby improving the safety, specificity, and tunability of next-generation cellular immunotherapies.</p>","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":"9 2","pages":"150-158"},"PeriodicalIF":4.8,"publicationDate":"2026-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13175979/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147964800","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Challenges and solutions for upstream processing of complex biologics.","authors":"Shuya Xu, Yunlong Wang, Yawen Yu, Felix Zhu, Ying Li","doi":"10.1093/abt/tbag008","DOIUrl":"10.1093/abt/tbag008","url":null,"abstract":"<p><p>Unlike conventional monoclonal antibodies, complex biologics-such as bispecific antibodies and fusion proteins-often face challenges including lower expression levels, higher mispairing rates, and greater sensitivity to culture conditions, which can collectively limit both titer and product quality. Leveraging process development case studies, this review systematically explores upstream bioprocessing strategies aimed at mitigating these challenges, with a focus on titer enhancement, lactate metabolism regulation, acidic charge variants control, reduction of aggregates and fragments, and glycosylation optimization. Finally, a perspective toward future upstream development strategies is discussed.</p>","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":"9 2","pages":"139-149"},"PeriodicalIF":4.8,"publicationDate":"2026-03-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13070570/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147677325","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Preventing physical and chemical degradation of the LABL-Fc-MOG_R5, a bifunctional peptide inhibitor, with formulation development approaches.","authors":"Lun Xin, Rucha Mahadik, Monika Prorok, Marcela Solis Rodriguez, Kai Qing Chan, Thomas Tolbert, Yunsong Li, Teruna J Siahaan","doi":"10.1093/abt/tbag005","DOIUrl":"10.1093/abt/tbag005","url":null,"abstract":"<p><strong>Background: </strong>LABL-Fc-MOG_R5 is a bifunctional peptide inhibitor designed to modulate immune responses in multiple sclerosis, including primary progressive multiple sclerosis. Although therapeutically promising, its development has been hindered by poor physical stability, particularly rapid precipitation under standard formulation conditions.</p><p><strong>Methods: </strong>A high-throughput, multi-phase formulation strategy was implemented to improve conformational and colloidal stability. An initial Phase 1 evaluation used intrinsic fluorescence differential scanning fluorometry and polyethylene glycol (PEG) solubility assays to screen 96 formulations spanning a range of pH values, buffer systems, and excipients. Then, a Phase 2 study applied a definitive screening design to evaluate key formulation variables-including buffer type, excipients, and surfactant concentration-at reduced protein concentration in 96-well plates. A final Phase 3 study compared the lead formulation from Phase 2 with the control formulation at higher protein concentration in glass vials.</p><p><strong>Results: </strong>The Phase 1 study revealed pronounced sensitivity of LABL-Fc-MOG_R5 to pH and ionic strength. Divalent anionic buffers (e.g. citrate, succinate) and excipients such as sucrose and hydroxypropyl-β-cyclodextrin substantially reduced aggregation propensity. In the Phase 2 study, an optimized formulation was identified-10 mM sodium acetate, pH 5.3, 125 mM sucrose, 150 mM HBP-LB-βCD, and 0.025% polysorbate 80-based on statistical modeling. This formulation demonstrated markedly improved resistance to aggregation, fragmentation, and subvisible particle formation under accelerated stress relative to the control.</p><p><strong>Conclusions: </strong>This work underscores the value of excipient selection and rational design of experiments in stabilizing complex fusion proteins. The optimized formulation provides a significantly enhanced stability profile and supports further development of LABL-Fc-MOG_R5 toward clinical evaluation for primary progressive multiple sclerosis.</p>","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":"9 3","pages":"215-229"},"PeriodicalIF":4.8,"publicationDate":"2026-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13317743/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148362811","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Durable disease control following treatment with ALMB-0168, a novel Cx43 hemichannel agonist monoclonal antibody, in femoral osteosarcoma with lung metastases after chemotherapy failure: a case report.","authors":"Xianbiao Xie, Yiying Bian, Xiugao Yang, Yanfeng Zhang, Junqiang Yin, Jingnan Shen","doi":"10.1093/abt/tbag007","DOIUrl":"10.1093/abt/tbag007","url":null,"abstract":"<p><p>Limited progress has been made over the past three decades in improving survival for patients with osteosarcoma after standard therapy. ALMB0168 is a first-in-class therapeutic antibody agonist targeting Cx43 hemichannels and has demonstrated to suppress the growth and migration of osteosarcoma in preclinical studies. This article presents a case of a patient with bilateral femoral osteosarcoma who was treated with ALMB0168. The patient, who had metastases in both contralateral limb and lung with ≥3 prior lines chemotherapy, showed immediate lesion shrinkage upon treatment and subsequently achieved durable partial response with a long duration of the response. The patient had a progression-free survival of 23 months to date, and the treatment was still ongoing. This case highlights that ALMB0168 may represent a promising treatment option for patients with metastatic or unresectable osteosarcoma after standard chemotherapy.</p>","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":"9 2","pages":"119-124"},"PeriodicalIF":4.8,"publicationDate":"2026-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13042229/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147610171","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analysis of the use of monoclonal antibodies in the treatment of Crohn's disease.","authors":"Alexander V Blagov, Marina D Sazonova, Anastasia I Ryzhkova, Vasily P Karagodin, Mikhail A Popov, Egor Yu Budnikov, Alessio L Ravani, Alexander N Orekhov, Margarita A Sazonova, Yuri V Arkhipenko","doi":"10.1093/abt/tbag006","DOIUrl":"10.1093/abt/tbag006","url":null,"abstract":"<p><p>Crohn's disease (CD) is a chronic inflammatory bowel disease with increasing global prevalence, significantly impacting patients' quality of life and healthcare costs. The introduction of monoclonal antibodies has revolutionized CD management, offering targeted therapy against specific inflammatory pathways. This review systematically analyzes the current state of monoclonal antibody therapy, including anti-TNF-α agents (infliximab, adalimumab, certolizumab pegol), anti-integrin antibodies (vedolizumab), and anti-cytokine therapies (ustekinumab, risankizumab). Despite remarkable therapeutic advances, significant limitations persist, including primary non-response (20%-40%), secondary loss of response (13%-20% annually), immunogenicity, safety concerns, and substantial economic burden. We propose evidence-based strategies to address these challenges, including therapeutic drug monitoring, combination therapy, and personalized medicine approaches. Furthermore, we identify promising novel therapeutic targets such as IL-36, IL-17C, SMAD7, TL1A, complement components, and microbiome-related factors. Targeting two or more specific targets simultaneously appears to be a promising direction of research for the development of bi- and polyspecific monoclonal antibodies capable of interfering with multiple pathological pathways in CD. The integration of advanced antibody engineering, personalized medicine, and innovative delivery systems represents the future direction for overcoming current limitations. Achieving sustained remission for all patients through safe, effective, and accessible therapeutic interventions remains the ultimate goal in CD management.</p>","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":"9 2","pages":"101-118"},"PeriodicalIF":4.8,"publicationDate":"2026-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13042251/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147610189","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction to: CAR NK cell therapy for solid tumors: potential and challenges.","authors":"","doi":"10.1093/abt/tbag004","DOIUrl":"https://doi.org/10.1093/abt/tbag004","url":null,"abstract":"<p><p>[This corrects the article DOI: 10.1093/abt/tbaf019.].</p>","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":"9 1","pages":"100"},"PeriodicalIF":4.5,"publicationDate":"2026-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12967325/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147390481","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A glycoengineered anti-ROR1 antibody, GE-zilovertamab, selectively enhances antibody-dependent cellular cytotoxicity against chronic lymphocytic leukemia.","authors":"Md Kamrul Hasan, George Widhopf Ii, Thomas J Kipps","doi":"10.1093/abt/tbag001","DOIUrl":"10.1093/abt/tbag001","url":null,"abstract":"<p><p>Receptor tyrosine kinase-like orphan receptor 1 (ROR1) is selectively expressed on chronic lymphocytic leukemia (CLL) B cells and certain cancers, but is absent from normal B cells and healthy adult tissues. GE-zilovertamab, an afucosylated anti-ROR1 IgG1 antibody, is engineered to increase FcγRIIIA binding and thereby enhance antibody-dependent cellular cytotoxicity (ADCC). Co-culture assays were performed using CLL cell lines (MEC1, MEC1-ROR1) and primary CLL cells with Jurkat-Lucia™ NFAT-CD16, NK, or peripheral blood mononuclear cell effectors. Treatments included the anti-CD20 mAb rituximab, anti-ROR1 mAbs (GE-zilovertamab, zilovertamab), and the endocytosis inhibitor prochlorperazine, and ADCC was quantified. GE-zilovertamab showed significantly higher ADCC than its parental antibody and activity that was comparable to that of rituximab. We find that the endocytosis inhibitor prochlorperazine further increased this effect. GE-zilovertamab is a promising next-generation immunotherapeutic for CLL, combining selective targeting of ROR1 with the potential to reduce therapy-induced immunodeficiency compared with anti-CD20 antibodies.</p>","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":"9 1","pages":"70-75"},"PeriodicalIF":4.8,"publicationDate":"2026-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12967326/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147391443","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}