Antibody Therapeutics最新文献

{"title":"DISCOVERY OF A CYNOMOLGUS MONKEY-CROSS-REACTIVE ANTI-HUMAN CD3 MAB FOR T CELL ENGAGERS","authors":"Qin Mei, George Wang, JieYing Liu, Yunying Chen, J. Gu, Siwei Nie","doi":"10.1093/abt/tbad014.004","DOIUrl":"https://doi.org/10.1093/abt/tbad014.004","url":null,"abstract":"Abstract Background Anti-CD3 based T cell engager antibodies can redirect cytotoxic activity of T cells in a non-MHC restricted fashion to kill tumor cells effectively. Therefore, the discovery of an anti-CD3 antibody capable of activating T cells in the presence of tumor cells is highly desirable. Recently, many anti-CD3 bispecific antibodies (bsAbs) entered clinical trials. Despite the promising efficacy of anti-CD3 bsAbs, safety issues arose and establishing a proper therapeutic window between efficacy and safety became a challenge. One of the safety concerns for anti-CD3 bsAbs is the cytokine release syndrome due to T-cell activation. Recent studies have shown that this safety challenge can be mitigated by selecting an anti-CD3 antibody with the appropriate binding epitope, CD3 affinity and binding kinetics (on and off rate). Methods by using WuXi Biologics’ state-of-the-art hybridoma platform, an anti-CD3 Ab was discovered through a combination of immunization and screening strategies. Results the selected anti-CD3 Ab demonstrates moderate affinity and fast-on fast-off binding kinetics against both human and cynomolgus CD3 molecules. Once constructed into T cell engagers (TCEs) using this anti-CD3 Ab with TAA binding arms in WuXiBody® format, the obtained TCEs mediated efficient anti-tumor activity, but induced low levels of cytokine production by T cells. Conclusions WuXi Biologics has discovered an anti-CD3 Ab with desired binding properties to human CD3. As shown in two showcases, the TCEs constructed using this anti-CD3 Ab can elicit efficient T cell cytotoxicity against tumor cells but low levels of cytokine release. The cross-reactivity of the anti-CD3 Ab enables preclinical assessments of toxicity in NHP.","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49649809","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":"Efficient production of bispecific antibodies-optimization of transfection strategy leads to high-level stable cell line generation of a Fabs-in-tandem immunoglobin.","authors":"Shiyong Gong,&nbsp;Chengbin Wu","doi":"10.1093/abt/tbad013","DOIUrl":"https://doi.org/10.1093/abt/tbad013","url":null,"abstract":"<p><p>Bispecific antibodies (bsAbs) are often composed of more than two component chains, such as Fabs-in-tandem immunoglobin (FIT-Ig) comprising three different component chains, which bring challenges for generating a high proportion of the correctly assembled bsAbs in a stable cell line. During the CHO-K1 stable cell line construction of a FIT-Ig, we investigated the FIT-Ig component chain ratio in transfection, where two sets of expression vectors were designed. Both designs utilized two vectors for co-transfection. Multiple transfections with plasmid ratio adjustment were applied, and the resultant minipools were evaluated for expression titer and quality of produced FIT-Ig. The results suggested that abundant outer Fab short chains (twofold chain genes versus other chains) can promote complete FIT-Ig assembly and therefore reduce the fragmental impurities of FIT-Ig. This adjustment of the component chain ratios at the beginning is beneficial to FIT-Ig stable cell line generation and brings favorable clones to process development.</p>","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":null,"pages":null},"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/88/39/tbad013.PMC10365153.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9878120","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: A mammalian cell display platform based on scFab transposition.","authors":"","doi":"10.1093/abt/tbad015","DOIUrl":"https://doi.org/10.1093/abt/tbad015","url":null,"abstract":"<p><p>[This corrects the article DOI: 10.1093/abt/tbad009.].</p>","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10365151/pdf/tbad015.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10229150","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":"JSKN003, A NOVEL BIPARATOPIC ANTI-HER2 ANTIBODY-DRUG CONJUGATE, EXHIBITS POTENT ANTITUMOR EFFICACY","authors":"Pilin Wang, K. Guo, Jianjian Peng, Jie Sun, Ting Xu","doi":"10.1093/abt/tbad014.009","DOIUrl":"https://doi.org/10.1093/abt/tbad014.009","url":null,"abstract":"Abstract In human advanced solid tumors, expression of HER2 protein has been reported in various tumor tissues and a variety of cultured tumor cell lines including breast cancer, gastric cancer, pancreatic cancer, lung cancer, colorectal cancer, and ovarian cancer. Due to the critical roles of HER2 in carcinogenesis, two main targeted therapies have been developed in the past two decades to block the HER2-driven pathways, which include small molecule compounds that inhibit the tyrosine kinase activity of the intracellular domain, and mono-antibodies (mAbs) that target the extracellular domain (ECD) of the receptors. JSKN003 is an antibody-drug conjugate (ADC) comprised of a recombinant, humanized anti-human epidermal growth factor receptor 2 (HER2) bispecific antibody conjugated to a topoisomerase I inhibitor via a dibenzocyclooctyne tetrapeptide linker. The anti-HER2 component, KN026, is a recombinant, humanized bispecific antibody that targets both extra-cellular domains II (pertuzumab binding site) and IV (trastuzumab binding site) of HER2. JSKN003 showed high affinity binding to human HER2 with KDs of 2.209 E-10M, which is comparable to its parental antibody KN026 and bound to NCI-N87 and BxPC-3 cells in a concentration-dependent manner. At the same time JSKN003 showed more extensive and faster internalization than DS8201 on NCI-N87 cells. As expected, JSKN003 showed directly inhibits growth by targeting HER2 positive tumor model (NCI-N87 and BT474 cell models). The single dose and multiple dose pharmacokinetics study in cynomolgus monkey indicated that JSKN003, total antibody and DXd had general linear dynamic characteristics, and pharmacokinetics parameters showed no significant differences between males and females in the range 0.3-30 mg/kg. The HNSTD (highest non-severely toxic dose) of JSKN003 was determined as 30mg/kg in cynomolgus monkeys. These preclinical data suggest that JSKN003 could potentially benefit patients with tumors co-expressing HER2 through improved drug selectivity and efficacy. JSKN003’s safety, tolerability and preliminary anti-tumor activity are currently being evaluated in a first-in-human phase I study in advanced stage solid tumors in Australia (NCT05494918) using a BOIN design. This ADC is targeted on the subjects who has HER2 expression and/or HER2-gene mutation and may address an unmet medical need for these patients.","PeriodicalId":36655,"journal":{"name":"Antibody Therapeutics","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45928177","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 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":null,"pages":null},"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":null,"pages":null},"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":null,"pages":null},"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":null,"pages":null},"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":null,"pages":null},"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":null,"pages":null},"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}