Antibody Therapeutics最新文献

{"title":"A NOVEL MSLN×4-1BB BISPECIFIC ANTIBODY FOR SOLID TUMOR","authors":"Liansheng Cheng, Dayan Zhang, Wenting Liu, Wei Zhou, Xiaoli Zeng, Qun Zhao, G. Shen","doi":"10.1093/abt/tbad014.003","DOIUrl":"https://doi.org/10.1093/abt/tbad014.003","url":null,"abstract":"Abstract Background Mesothelin (MSLN) is a 70 KD glycosylphosphatidylinositol (GPI)-anchored cell surface glycoprotein that is rarely expressed in normal tissues but overexpressed in a variety of cancers, including mesothelioma, ovarian cancer, pancreatic cancer and breast cancer et.al. 4-1BB is a member of the tumor necrosis factor receptor superfamily that functions as a co-stimulatory molecule. Agonistic antibodies bind to 4-1BB, triggering a signaling cascade leading to T-cell activation and expansion of cytotoxic CD8+ T lymphocytes. Here, we developed two bispecific antibodies (bsAbs) targeting both MSLN and 4-1BB with an intact Fc fragment from human IgG1 or IgG4, named HK013-G1 and HK013-G4 respectively. We suspected that HK013-G1 can simultaneously exert the cytotoxic effect of CD8+T cells and NK cells on tumor cells expressing MSLN to achieve better antitumor efficacy. Methods Both HK013-G1 and HK013-G4 were constructed by fused a single-chain variable fragment (scFv) targeting hu4-1BB to the C terminus of an anti-MSLA nanobody. And their affinity was optimized to making it highly effective in tumor localization. Next, we tested the killing ability of bsAbs-mediated PBMC or NK92 against tumor cells with different expression levels of MSLN in vitro. And the IFN-γ secretion was detected when CD8+T cells co-cultured with MSLN+ or MSLN- cells in the presence of antibodies. Also, the 4-1BB agonist activity of bsAbs was measured in a luciferase report gene assay. To confirm the safety of HK013-G1, non-specific activation of 4-1BB signal mediated by Fc receptor and CRS was evaluated in vitro. Finally, we compared the antitumor activity of two bispecific antibodies in both MC38/hMSLN and CT26/hMSLN tumor model and hepatotoxicity as well as cardiotoxicity was evaluated. Results Affinity-optimized HK013-G1 has an order of magnitude greater affinity for MSLN(KD≈10−9M) than 4-1BB (KD≈10−8M). HK013-G1 induced stronger PBMC against tumor cells than MOARb009 while HK013-G4 does not. Also, HK013-G1 could only mediate the killing of NK92 on MSLN-positive tumor cells. In co-cultured assay, HK013-G1 had superior ability to stimulate CD8+T cell secretion of IFN-γ than urelumab in the presence of MSLN. In luciferase reporter assay, the bsAbs-induced 4-1BB activation is dependent on expression level of MSLN. In addition, HK013-G1 was shown no stronger ability to inducing non-specific activation of 4-1BB signal mediated by Fc receptor and CRS in vitro. Compared with HK013-G4, HK013-G1 showed a more significant anti-tumor effect in both MC38/hMSLN and CT26/hMSLN tumor model. And, HK013-G4 showed significant hepatotoxicity in mice while HK013-G1 not. Moreover, HK013-G1 can protect mice against tumor re-challenge. Conclusions HK013-G1, an MSLN×4-1BB bsAb with human IgG1 Fc fragment, prevents tumor development by killing tumor cells directly via effector functions mediated by NK and cytotoxic T cells. More importantly, HK013-G1 showed no stronger toxic side effect","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46993982","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"ENHANCED KINETICS ANALYSIS OF PROTEINS AND LARGE BIOMOLECULES USING NOVEL HIGH SENSITIVITY PROBE","authors":"John Zhang, Weijing Gu, Hongshan Li, Pu Li","doi":"10.1093/abt/tbad014.026","DOIUrl":"https://doi.org/10.1093/abt/tbad014.026","url":null,"abstract":"Abstract Introduction Bio-layer interferometry (BLI) has gained significant interest as a label-free technique for the detection and kinetic analysis of diverse biomolecules such as antibodies, proteins, and small molecules. The technology relies on the phase shift-wavelength correlation generated between interference patterns at the tip of the biosensor probe where molecules associate and dissociate. However, current biosensors face challenges regarding sensitivity with small molecules/peptides and compatibility with large biomolecules like nanomaterials. Traditional BLI often produces inverted signals when nanomaterials bind which hinders accurate kinetics analysis. Overcoming these limitations is crucial for expanding the range of applications and enhancing the performance of BLI-based detection systems. Significance In this study, we have developed an improved BLI sensor, Gator® SA XT, which features newly designed streptavidin-based surface capable of loading biotinylated ligands as small as 1.5 kDa. Compared to traditional BLI streptavidin probes, the SA XT probes exhibit a 3-5 times higher signal intensity. Moreover, the incorporation of a novel optical coating layer enables the detection of large biomolecules such as lipid nanoparticles without signal inversion. This advancement in biosensor technology facilitates the detection of ligands and their analytes at lower concentrations and expands the range of compatible analytes for BLI-based applications. Methods To enhance the sensitivity of the interference patterns, we utilized a proprietary optical coating layer with a refractive index significantly lower than that of proteins and other biomolecules. We assessed the sensitivity and sensing distance of the optical coating layer using a layer-by-layer model system. Binding cycles of biotinylated protein A and human IgG were repeated until the theoretical biolayer thickness reached approximately 700 nm. Results Comparative analysis of binding signals between the newly designed SA XT probes and traditional SA probes were conducted for various biomolecules. The SA XT probes demonstrated significantly higher binding signals for oligos (2.8-fold), peptides (3.0-fold), Protein A (4.1-fold), PDL1 (4.5-fold), and IgG (4.3-fold). Furthermore, the unique optical properties of the SA XT probes prevented signal inversion enabling the detection of biomolecules as large as 2 MDa. Using a layer-by-layer model system, the SA XT probes successfully detected a biolayer thickness of 700nm without signal inversion. Additionally, we demonstrated the detection of lipid nanoparticles and subsequent biomolecule bindings using the SA XT probes. Conclusions In conclusion, we have designed a novel biosensor for BLI that enables the detection of a wider range of biomolecules with high sensitivity. The SA XT probes, coupled with the proprietary optical coating layer, have overcome the limitations of traditional BLI probes and facilitated the generation of reliable and","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43814439","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A NOVEL IMMUNOSTIMULATORY PD-L1/OX40 TETRAVALENT BISPECIFIC ANTIBODY FOR CANCER IMMUNOTHERAPY","authors":"Baocun Li, Xuan Wu, Shiyong Gong, Zhou Lv, Nianying Zhang, Yu Zhang, G. Naren, Danqing Wu, Jianfu Wu, Fan Liu, Rui Zhang, Chengbin Wu","doi":"10.1093/abt/tbad014.008","DOIUrl":"https://doi.org/10.1093/abt/tbad014.008","url":null,"abstract":"Abstract Single agent immune checkpoint therapy has shown substantial and durable clinical activity in many tumor types; however, only a fraction of the patients could benefit from this approach. To improve beyond the anti-PD-1/PD-L1 treatment options, bispecific antibodies (BsAb) that combines PD-L1 blockade and conditional co-stimulatory receptor activation simultaneously in one molecule have been developed and demonstrated superior anti-tumor activity in pre-clinical models. However, many of these PD-L1 based BsAb faced challenge in clinical development due to insufficient activity or unexpected toxicity. Here, we demonstrated that OX40 might be a more suitable partner for PD-L1 based BsAb design than other agonistic targets (CD27 and 4-1BB, etc.) currently in clinical studies. A novel Fc silenced tetravalent PD-L1/OX40 (EMB-09) BsAb targeting optimal OX40 binding epitope has been developed based on EpimAb’s proprietary FIT-Ig® technology. Results showed that EMB-09 maintained the parental mAb binding characteristic and retained the functional properties of each parental mAb including OX40 agonistic as well as PD-L1/PD1 inhibitory pathways. In addition, EMB-09 induced OX40 activation only in the context of PD-L1 engagement. Concurrent PD-L1/PD-1 blockade and OX40 co-stimulation by EMB-09 led to synergistic activation of T cell in vitro and exerted superior anti-tumor activity in mouse tumor models compared to anti-PD-L1 mAb. The underlining mechanism was extensively analyzed, which indicated an increased CD8+ tumor-infiltrating T-cells (TIL) as well as enhanced CD8 TIL activation status upon EMB-09 treatment. Additionally, EMB-09 was well tolerated in cynomolgus monkeys at high dose levels with a favorable safety and PK profile in a GLP-TOX study. In conclusion, as a PD-L1/OX40 BsAb with a novel biology mechanism, EMB-09 demonstrated a markedly improved anti-tumor activity compared to anti-PD-L1 mAb. The first-in-human clinal study of EMB-09 has been initiated (NCT05263180).","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47312189","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"ANTIBODY CO-FORMULATION TOOLBOX AND CAPABILITIES ESTABLISHED IN WUXI BIOLOGICS","authors":"Anyuan Liu, J. Weng, Fangyuan Zhou, Kewei Wang, Hongbing Wu, S. Wang, Jeremy Guo","doi":"10.1093/abt/tbad014.018","DOIUrl":"https://doi.org/10.1093/abt/tbad014.018","url":null,"abstract":"Abstract Introduction Co-formulation containing two or more antibodies (mAbs) is deemed to hold distinct merits such as better treatment efficacy, higher efficiency and extended intellectual property right, attracting the demands from both patients and pharmaceutical companies. However, there are only limited numbers of approved drug products, partially due to the technical challenges in formulation development and analytical methods. Herein, we present WuXi Biologics efforts to accelerate the development of co-formulation drug products. Methodology We have established an antibody co-formulation specific toolbox with a dedicated team for co-formulation product development addressing the formulation and analytical challenges. Our co-formulation analytical expertise includes size exclusion-high performance liquid chromatography (SE-HPLC), caliper-sodium dodecyl sulfate reduced and non-reduced (Caliper-SDS-R & NR), imaged capillary isoelectric focusing (iCIEF), ion-exchange chromatography (IEC), reverse phase-liquid chromatography (RP-LC), hydrophobic interaction chromatography (HIC), Composition-Gradient Multi-Angle Light Scattering (CG-MALS), differential scanning calorimetry (DSC), enzyme-linked immunosorbent assay (ELISA) based and/or cell based potency, and peptide mapping with mass spectrometry etc. Results Remarkably, couple of antibodies co-formulation cases have completed successfully. Take one co-formulation case for instance, specifically, no substantial molecular interactions were observed between the two antibodies according to the results of differential light scattering (DLS) and CG-MALS. Besides, the main peaks of two mAbs were co-eluted in SE-HPLC and Caliper-SDS-NR, respectively. SE-HPLC and Caliper-SDS-NR methods were optimized to evaluate the purity of this co-formulation. As a result, the purity of these two mAbs in this co-formulation was comparable with its individual antibody respectively. Given the isoelectric point of two mAbs in this co-formulation differs by 1.0, iCIEF has been developed to separate the peaks of two mAbs completely. In another case, the isoelectric point of two mAbs differed by just 0.4, CEX has been developed. iCIEF and CEX method were optimized to evaluate the charge variants and determine the concentration ratio of these two mAbs in co-formulation product.","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48975339","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"OVERCOMING TECHNICAL CHALLENGES OF BISPECIFIC AND MULTI-SPECIFIC ANTIBODIES USING NOVEL TECHNOLOGY PLATFORMS","authors":"Jianqing Xu, Shuang Wang, Xinzhao Fan, George Wang, J. Gu, Siwei Nie","doi":"10.1093/abt/tbad014.007","DOIUrl":"https://doi.org/10.1093/abt/tbad014.007","url":null,"abstract":"Abstract Background Bispecific antibodies (bsAbs) and multispecific antibodies (msAbs) are a growing class of next generation biotherapeutics. However, multiple challenges, such as chain mispairing and developability issues are often associated with these complex modalities. Methods WuXi Biologics has established bsAb and msAb engineering and development platforms. By replacing CH1/CL domains of one antibody Fab by corresponding T cell receptor (TCR) Cβ/Cα domains, WuXiBody® bsAb technology can ensure the correct heavy-light chain pairing. WuXi Biologics has developed the SDArBody™ platform, which utilizes single domain antibodies (VHH) as building blocks to facilitate the exploration of more complex biologics, such as msAbs. Results as shown in case studies, the WuXiBody® technology is able to assemble regular mAbs in a ‘plug-and-play’ manner, and that the SDArBody® can be utilized to identify VHH leads and then assemble into msAbs. Both platforms can generate highly functional bsAb and msAb with good developability. Conclusions The establishment of WuXiBody® and SDArBody® platforms could greatly meet the needs of biologics developers in pursuit of different biology and therapeutic approaches through bsAbs and msAbs.","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48231724","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The influence of variable-heavy chain families on IgG₂, ₃, ₄, FcγRs and B-cell superantigens protein G and L binding using biolayer interferometry.","authors":"Anthony M Deacy,&nbsp;Samuel Ken-En Gan","doi":"10.1093/abt/tbad016","DOIUrl":"https://doi.org/10.1093/abt/tbad016","url":null,"abstract":"<p><p>As the most abundant immunoglobulin in blood and the most common human isotype used for therapeutic monoclonal antibodies, the engagement and activation of its Fc receptors by IgGs are crucial for antibody function. Assumed to be relatively constant within subtypes, recent studies reveal that antibody variable regions exert distal effects of modulating antibody-receptor interactions on antibody isotypes. These variable (V)-region distal effects are also expected for the IgG subtypes. With an in-depth understanding of the V-region effects, researchers can make a more informed antibody engineering approach and antibody purification strategy accounting for the functions of microbial immune evasion . In this study, we created a panel of IgG₂/IgG₃/IgG₄ antibodies by changing the V_H family (V_H1-7) frameworks while retaining the complementary determining regions of pertumuzab and measured their interactions with FcγRIa, FcγRIIa_H167, FcγRIIa_R167, FcγRIIb/c, FcγRIIIa_F176, FcγRIIIa_V176, FcγRIIIb_NA1 and FcγRIIIb_NA2 receptors alongside B-cell superantigens Protein L and G using biolayer interferometry. The panel of 21 IgGs demonstrated that the V_H frameworks influenced receptor binding sites on the constant region in a non-canonical manner. However, there was minimal influence on the binding of bacterial B-cell superantigens Proteins L and Protein G on the IgGs, showing their robustness against V-region effects. These results demonstrate the role of V-regions during the humanization of therapeutic antibodies that can influence FcR-dependent immune responses while retaining binding by bacterial B-cell superantigens for antibody purification. These <i>in vitro</i> measurements provide a clue to detailed antibody engineering and understanding of antibody superantigen functions that would be relevant with <i>in vivo</i> validation.</p>","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":"6 3","pages":"182-193"},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/88/28/tbad016.PMC10481891.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10177231","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":"FULLY AI-DRIVEN HUMANOID VHH PHAGE LIBRARY","authors":"Yangyang Zhao, Le Niu, Xuemin Pan, Xingda Ye, Quan Yu, Yupeng Zhu, Yile Chen, Zhiwu Sun, Yunfei Long, Yi Li","doi":"10.1093/abt/tbad014.020","DOIUrl":"https://doi.org/10.1093/abt/tbad014.020","url":null,"abstract":"Abstract Background & Significance VHHs are small and stable fragments that have great potential as therapeutics due to their small size, stability, versatility, and potential for oral administration. The traditional source of VHHs is camelids, but humanization is usually needed for therapeutic development. A human VHH library is highly desirable for the generation of therapeutic VHHs, but natural human VH domains are usually unstable as standalone units. We developed a humanoid VHH library of AI-designed sequences that both resemble camelid VHHs in terms of stability and have such high human content that humanization is no longer needed. Methods In this study, we present a fully AI-driven approach for the de novo design of a VHH phage library. Firstly, public camelid data and nearly one million private human sequences were collected. Secondly, one autoregressive AI model was trained on human data and another AI model was trained on the mixed data of humans and camels. Thirdly, the CDR1, CDR2, CDR3 regions of VHH were all generated by the mentioned two AI models. Finally, an ultra large quantity (4E10) of VHH sequences generated by AI were utilized to build the Humanoid VHH phage library. Results In order to verify the effectiveness of our method, we randomly synthesized and expressed 26 VHH antibodies from our AI based library. At the same time, 3 human VH molecules reported in previous literature were included as positive controls. First of all, the success rate of expression is 96.1%, which is much higher than 72% of Progen and 66% of ESMdesign. Secondly, the average titer is 59.6mg/L, which is 1.5 times the average value of the positive control group. Thirdly, the hydrophobicity of 80% de novo sequences is comparable to the positive control group. Moreover, the immunogenicity of all AI sequences is less than the average value of the positive control group according to our proprietary algorithms. Finally, the diversity and naturalness of the Humanoid VHH phage library are also excellent. Conclusions In conclusion, we have developed a fully AI-driven solution that could stably and massively generate human-like VHH sequences satisfying multiple requirements (including high titer, low hydrophobicity, low immunogenicity and ultra high success rate of expression, high diversity, high naturalness) simultaneously. As VHH is a powerful therapeutic fragment, our approach has the potential to accelerate nanobody and bispecific antibody drug development.","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46529624","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction: AB-Amy: machine learning aided amyloidogenic risk prediction of therapeutic antibody light chains.","authors":"","doi":"10.1093/abt/tbad012","DOIUrl":"https://doi.org/10.1093/abt/tbad012","url":null,"abstract":"<p><p>[This corrects the article DOI: 10.1093/abt/tbad007.].</p>","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":"6 3","pages":"180"},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/0a/64/tbad012.PMC10365152.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10229151","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":"TARGETING TREG CELLS BY TNFR2 ANTIBODY INDUCES TUMOR REGRESSION IN VIVO","authors":"Zuoan Yi, C. He, Yueh-Mei Hso, M. Strainic, Yong Yin, W. Zhai","doi":"10.1093/abt/tbad014.015","DOIUrl":"https://doi.org/10.1093/abt/tbad014.015","url":null,"abstract":"Abstract Background Tumor necrosis factor receptor 2 (TNFR2) is considered an appealing target due to its low-level expression on immune cells, but it could be upregulated on regulatory T cells (Tregs) in the tumor microenvironment which plays key roles in Treg proliferation and function. It has been demonstrated that Tregs with high TNFR2 expression were the most suppressive subsets among all Treg populations in the tumor. While early studies showed that TNFR2 co-stimulates naïve T cells, it has been revealed later that TNFR2 also limits CD8+ T-cell-mediated viral clearance and anti-tumor immunity by inducing rapid contraction of CD8+ T cells. Consistent with these findings, several publications reported that anti-TNFR2 antibody treatment exhibited robust antitumor efficacy. In short, TNFR2 signaling is critical in regulating immune response in different diseases. In the current study, the anti-human TNFR2 antibody SBT-1901 was developed and the anti-tumor activity was evaluated. Materials and methods SBT-1901 was generated through hybridoma and humanization technologies. The binding affinity and specificity were tested by ELISA, FACS, and OCTET. The function of SBT-1901 was tested in TNFR2-Fas overexpressing Ramos cells and in Treg proliferation assay. The in vivo anti-tumor activity and pharmacokinetics of SBT-1901 were evaluated in the human TNFR2 transgenic mouse model (Biocytogen) bearing MC38 tumor. Results SBT-1901 binds to the extracellular domain of human and cynomolgus TNFR2 with the affinity of single digit-nanomolar. It competes with TNFα for binding to TNFR2 receptor and inhibits TNFα-induced TNFR2-Fas overexpressing RAMOS cell death. SBT-1901 also blocks TNFα induced Treg proliferation in human PBMC. In addition, SBT-1901 significantly inhibits MC38 tumor growth in a dose-dependent manner as a monotherapy and enhances anti-tumor efficacy of mPD-1 antibody in a combination study in human TNFR2 transgenic mouse model. SBT-1901 is currently under preclinical development. Conclusions Our studies show that SBT-1901 exhibits a very potent anti-tumor efficacy in vivo as a single agent or the combination. Therefore, it is highly valuable to further develop SBT-1901 for human cancer treatment.","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43558052","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"APPLICATION OF EXCHABODY TECHNOLOGY FOR PAIRING VHHS TO ACHIEVE SYNERGISTIC EFFECTS","authors":"Yi Luo, Xiaoxiao Zhan, Yilong Shen, Ziyang Sheng, Yin Zhu, Mingyue Huang","doi":"10.1093/abt/tbad014.025","DOIUrl":"https://doi.org/10.1093/abt/tbad014.025","url":null,"abstract":"Abstract Objective Single-domain antibodies, such as VHH and nanobody, have shown potential for use in therapy and diagnostics. One application of VHHs is the tethering of two fragments to different epitopes on the same target, which is difficult to achieve with conventional antibodies. Synergistic heterologous VHH dimers have higher affinity, better specificity, and broad applications in developing high-affinity monoclonal antibodies, bispecific antibodies, ADCs, and CAR-Ts. However, finding the best pair of VHHs for these applications requires combinational screening, which is traditionally a time-consuming and costly process. The objective of this study is to develop a technology that can quickly screen and pair two synergistic VHHs without the need to express tandem VHH dimers. Methods The researchers developed proprietary tags and specific dockers that, when stabilized on a solid station, can capture any VHHs that the dockers recognize and pull them together to form a non-covalent dimer. This platform is called ExchaBody technology, and the VHH dimers formed this way are ExchaBodies. The researchers used this technology to conduct a bi-epitope screening campaign, where VHHs were first expressed as monomers with tags and then binned and grouped into different categories. VHHs were then paired with all reasonable combinations using ExchaBody technology, and these ExchaBodies were evaluated for their combined activities. Results ExchaBody technology was able to link any two VHHs together within one hour, and the resulting ExchaBodies had bivalent or bifunctional VHH activities. The bi-epitope VHH screening campaign, which would have taken months to complete using traditional methods, was finished within two weeks using ExchaBody technology, saving time and cost. The researchers were able to construct two lead molecules, a bi-specific VHH-Fc fusion protein and a tri-valent VHH molecule, using ExchaBody technology. These lead molecules were found to be superior to their counterparts on the market based on affinity and functional assays. Conclusion ExchaBody technology is a bispecific VHH screening and pairing platform that can quickly and cost-effectively create non-covalent, bispecific VHHs (ExchaBodies) without the need to express them. ExchaBodies possess the binding and cellular activities of a covalently linked, bispecific, tandem VHH dimer. This technology has broad applications in developing high-affinity monoclonal antibodies, bispecific antibodies, ADCs, and CAR-Ts.","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43436208","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}