Monoclonal Antibodies in Immunodiagnosis and Immunotherapy最新文献_第2页

Recombinant Antibody-Producing Stable CHOK1 Pool Stability Study. 重组抗体产生稳定 CHOK1 池稳定性研究。

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Pub Date : 2024-08-01 Epub Date: 2024-07-22 DOI: 10.1089/mab.2024.0008

Bailin Tu, Zhihong Lin, Jeff Moore, Archana Krishnan Sekaran, Miranda J Wesley, De Yu Mao, Mark Gibson, Wan-Ching Lai, John Boggs, Thomas Slowik, Yeni Natalia C Perez-Gelvez, Ryan Bonn, Tracey Rae, Jeremy Minshull, Ferenc Boldog, Varsha Sitaraman, Scott Muerhoff, Philip Hemken

{"title":"Recombinant Antibody-Producing Stable CHOK1 Pool Stability Study.","authors":"Bailin Tu, Zhihong Lin, Jeff Moore, Archana Krishnan Sekaran, Miranda J Wesley, De Yu Mao, Mark Gibson, Wan-Ching Lai, John Boggs, Thomas Slowik, Yeni Natalia C Perez-Gelvez, Ryan Bonn, Tracey Rae, Jeremy Minshull, Ferenc Boldog, Varsha Sitaraman, Scott Muerhoff, Philip Hemken","doi":"10.1089/mab.2024.0008","DOIUrl":"10.1089/mab.2024.0008","url":null,"abstract":"Mammalian cell line stability is an important consideration when establishing a biologics manufacturing process in the biopharmaceutical and in vitro diagnostics (IVD) industries. Traditional Chinese hamster ovary (CHO) cell line development methods use a random integration approach that requires transfection, selection, optional amplification, screenings, and single-cell cloning to select clones with acceptable productivity, product quality, and genetic stability. Site-specific integration reduces these disadvantages, and new technologies have been developed to mitigate risks associated with genetic instability. In this study, we applied the Leap-In® transposase-mediated expression system from ATUM to generate stable CHOK1 pools for the production of four recombinant antibody reagents for IVD immunoassays. CHO cell line stability is defined by consistent antibody production over time. Three of the CHOK1 pools maintained productivity suitable for manufacturing, with high antibody yields. The productivity of the remaining CHOK1 pool decreased over time; however, derivative clones showed acceptable stability. l-glutamine had variable effects on CHOK1 cell line or stable pool stability and significantly affected antibody product titer. Compared with traditional random integration methods, the ATUM Leap-In system can reduce the time needed to develop new immunoassays by using semi site-specific integration to generate high-yield stable pools that meet manufacturing stability requirements.","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":" ","pages":"119-126"},"PeriodicalIF":0.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141735648","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}

引用次数: 0

Epitope Mapping of an Anti-Mouse CCR8 Monoclonal Antibody C₈Mab-2 Using Flow Cytometry. 使用流式细胞术绘制抗小鼠 CCR8 单克隆抗体 C8Mab-2 的表位图。

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Pub Date : 2024-08-01 Epub Date: 2024-06-05 DOI: 10.1089/mab.2024.0002

Hiyori Kobayashi, Hiroyuki Suzuki, Tomohiro Tanaka, Mika K Kaneko, Yukinari Kato

{"title":"Epitope Mapping of an Anti-Mouse CCR8 Monoclonal Antibody C8Mab-2 Using Flow Cytometry.","authors":"Hiyori Kobayashi, Hiroyuki Suzuki, Tomohiro Tanaka, Mika K Kaneko, Yukinari Kato","doi":"10.1089/mab.2024.0002","DOIUrl":"10.1089/mab.2024.0002","url":null,"abstract":"The C-C motif chemokine receptor 8 (CCR8) is highly and selectively expressed in regulatory T (Treg) cells and is associated with tumor progression. The massive accumulation of Treg cells into tumors suppresses the effector function of CD8+ cells against tumor cells. Therefore, selective depletion of Treg cells using anti-CCR8 monoclonal antibodies (mAbs) reinvigorates antitumor immune responses and improves responses to cancer immunotherapy. Previously, we developed an anti-mouse CCR8 (mCCR8) mAb, C8Mab-2, using the Cell-Based Immunization and Screening method. In this study, the binding epitope of C8Mab-2 was investigated using flow cytometry. The mCCR8 extracellular domain-substituted mutant analysis showed that C8Mab-2 recognizes the N-terminal region (1-33 amino acids) of mCCR8. Next, 1×alanine (or glycine) scanning and 2×alanine (or glycine) scanning were conducted in the N-terminal region. The results revealed that the 17-DFFTAP-22 sequence is important for the recognition by C8Mab-2, and Thr20 is a central amino acid of the epitope. These results revealed the involvement of the N-terminus of mCCR8 in the recognition by C8Mab-2.","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":" ","pages":"101-107"},"PeriodicalIF":0.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141248953","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}

引用次数: 0

Development of a Sensitive Anti-Mouse CCR5 Monoclonal Antibody for Flow Cytometry. 为流式细胞仪开发灵敏的抗小鼠 CCR5 单克隆抗体。

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Pub Date : 2024-08-01 Epub Date: 2024-06-05 DOI: 10.1089/mab.2024.0004

Hiroyuki Suzuki, Tomohiro Tanaka, Guanjie Li, Tsunenori Ouchida, Mika K Kaneko, Yukinari Kato

引用次数: 0

Development of Sensitive Anti-Mouse CCR5 Monoclonal Antibodies Using the N-Terminal Peptide Immunization. 利用 N 端肽免疫技术开发灵敏的抗小鼠 CCR5 单克隆抗体

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Pub Date : 2024-08-01 Epub Date: 2024-06-13 DOI: 10.1089/mab.2024.0009

Rena Ubukata, Hiroyuki Suzuki, Tomohiro Tanaka, Guanjie Li, Mika K Kaneko, Yukinari Kato

{"title":"Development of Sensitive Anti-Mouse CCR5 Monoclonal Antibodies Using the N-Terminal Peptide Immunization.","authors":"Rena Ubukata, Hiroyuki Suzuki, Tomohiro Tanaka, Guanjie Li, Mika K Kaneko, Yukinari Kato","doi":"10.1089/mab.2024.0009","DOIUrl":"10.1089/mab.2024.0009","url":null,"abstract":"One of the G protein-coupled receptors, C-C chemokine receptor 5 (CCR5), is an important regulator for the activation of T and B lymphocytes, dendritic cells, natural killer cells, and macrophages. Upon binding to its ligands, CCR5 activates downstream signaling, which is an important regulator in the innate and adaptive immune response through the promotion of lymphocyte migration and the secretion of proinflammatory cytokines. Anti-CCR5 monoclonal antibodies (mAbs) have been developed and evaluated in clinical trials for tumors and inflammatory diseases. In this study, we developed novel mAbs for mouse CCR5 (mCCR5) using the N-terminal peptide immunization. Among the established anti-mCCR5 mAbs, C5Mab-4 (rat IgG2a, kappa) and C5Mab-8 (rat IgG1, kappa), recognized mCCR5-overexpressing Chinese hamster ovary-K1 (CHO/mCCR5) and an endogenously mCCR5-expressing cell line (L1210) by flow cytometry. The dissociation constant (KD) values of C5Mab-4 and C5Mab-8 for CHO/mCCR5 were determined as 3.5 × 10-8 M and 7.3 × 10-9 M, respectively. Furthermore, both C5Mab-4 and C5Mab-8 could detect mCCR5 by western blotting. These results indicated that C5Mab-4 and C5Mab-8 are useful for detecting mCCR5 by flow cytometry and western blotting and provide a possibility to obtain the proof of concept in preclinical studies.","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":" ","pages":"112-118"},"PeriodicalIF":0.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141312337","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}

引用次数: 0

Cx₃Mab-4: A Novel Anti-Mouse CXCR3 Monoclonal Antibody for Flow Cytometry. Cx3Mab-4：用于流式细胞仪的新型抗小鼠 CXCR3 单克隆抗体。

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Pub Date : 2024-06-01 Epub Date: 2024-03-20 DOI: 10.1089/mab.2023.0024

Tsunenori Ouchida, Yu Isoda, Tomohiro Tanaka, Mika K Kaneko, Hiroyuki Suzuki, Yukinari Kato

{"title":"Cx3Mab-4: A Novel Anti-Mouse CXCR3 Monoclonal Antibody for Flow Cytometry.","authors":"Tsunenori Ouchida, Yu Isoda, Tomohiro Tanaka, Mika K Kaneko, Hiroyuki Suzuki, Yukinari Kato","doi":"10.1089/mab.2023.0024","DOIUrl":"10.1089/mab.2023.0024","url":null,"abstract":"C-X-C motif chemokine receptor 3 (CXCR3, CD183) is a G-protein-coupled receptor for CXCL9, CXCL10, and CXCL11. CXCR3 induces chemotaxis of immune cells and promotes inflammation. Various mouse models have been developed to mimic the pathogenesis of diseases and used in the evaluation of therapeutics for these diseases. Although CXCR3 is an attractive target to suppress inflammation, anti-CXCR3 therapeutic agents have not been approved. In this study, we established a novel anti-mouse CXCR3 (mCXCR3) monoclonal antibody, Cx3Mab-4 (rat IgG1, kappa), using the Cell-Based Immunization and Screening method. Flow cytometric analysis demonstrated that Cx3Mab-4 bound to mCXCR3-overexpressed Chinese hamster ovary-K1 (CHO/mCXCR3) cells, but did not react to parental CHO-K1 cells. The dissociation constant of Cx3Mab-4 was determined as 1.3 × 10-9 M, indicating that Cx3Mab-4 possesses a high affinity to mCXCR3-expressing cells. Cx3Mab-4 could be useful for targeting CXCR3-expressing cells in preclinical mouse models.","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":" ","pages":"90-95"},"PeriodicalIF":0.0,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140177676","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}

引用次数: 0

Charting the Course for Monoclonal Antibodies in Immunodiagnosis and Immunotherapy. 为免疫诊断和免疫疗法中的单克隆抗体绘制蓝图。

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Pub Date : 2024-06-01 Epub Date: 2024-06-10 DOI: 10.1089/mab.2024.0012

Cory L Brooks, Andrei Moroz

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Epitope Mapping of an Anti-CD44v4 Monoclonal Antibody (C₄₄Mab-108) Using Enzyme-Linked Immunosorbent Assay. 利用酶联免疫吸附试验绘制抗 CD44v4 单克隆抗体（C44Mab-108）的表位图。

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Pub Date : 2024-06-01 Epub Date: 2024-03-20 DOI: 10.1089/mab.2023.0022

Hiroyuki Suzuki, Mayuki Tawara, Aoi Hirayama, Nohara Goto, Tomohiro Tanaka, Mika K Kaneko, Yukinari Kato

{"title":"Epitope Mapping of an Anti-CD44v4 Monoclonal Antibody (C44Mab-108) Using Enzyme-Linked Immunosorbent Assay.","authors":"Hiroyuki Suzuki, Mayuki Tawara, Aoi Hirayama, Nohara Goto, Tomohiro Tanaka, Mika K Kaneko, Yukinari Kato","doi":"10.1089/mab.2023.0022","DOIUrl":"10.1089/mab.2023.0022","url":null,"abstract":"CD44 is a type I transmembrane glycoprotein and possesses various isoforms which are largely classified into CD44 standard (CD44s) and CD44 variant (CD44v) isoforms. Some variant-encoded regions play critical roles in tumor progression. However, the function of CD44 variant 4 (CD44v4)-encoded region has not been fully understood. Using peptide immunization, we developed an anti-CD44v4 monoclonal antibody, C44Mab-108, which is useful for flow cytometry, western blotting, and immunohistochemistry. In this study, we determined the critical epitope of C44Mab-108 by enzyme-linked immunosorbent assay (ELISA). We used the alanine (or glycine)-substituted peptides of the CD44v4-encoded region (amino acids 271-290 of human CD44v3-10) and found that C44Mab-108 did not recognize the alanine-substituted peptides of D280A and W281A. Furthermore, these peptides could not inhibit the recognition of C44Mab-108 in flow cytometry and immunohistochemistry. The results indicate that the critical binding epitope of C44Mab-108 includes Asp280 and Trp281 of CD44v3-10.","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":" ","pages":"85-89"},"PeriodicalIF":0.0,"publicationDate":"2024-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140177677","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}

引用次数: 0

Correction to: Establishment of a Novel Cancer-Specific Anti-HER2 Monoclonal Antibody H2Mab-250/H2CasMab-2 for Breast Cancers'' by Kaneko et al. Monoclonal Antibodies in Immunodiagnosis and Immunotherapy 2024;43(2):35-43; doi: 10.1089/mab.2023.0033. 更正：建立新型癌症特异性抗 HER2 单克隆抗体 H2Mab-250/H2CasMab-2 治疗乳腺癌''，Kaneko 等著，《免疫诊断和免疫疗法中的单克隆抗体》，2024;43(2):35-43; doi: 10.1089/mab.2023.0033。

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Pub Date : 2024-04-16 DOI: 10.1089/mab.2023.0033.correx

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The Gift That Keeps on Giving. 送人玫瑰，手有余香。

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Pub Date : 2024-04-01 DOI: 10.1089/mab.2024.29018.editorial

Thomas Kieber-Emmons

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Epitope Mapping of an Anti-Mouse CD39 Monoclonal Antibody Using PA Scanning and RIEDL Scanning. 利用 PA 扫描和 RIEDL 扫描绘制抗小鼠 CD39 单克隆抗体的表位图。

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Pub Date : 2024-04-01 Epub Date: 2024-03-20 DOI: 10.1089/mab.2023.0029

Yuki Okada, Hiroyuki Suzuki, Tomohiro Tanaka, Mika K Kaneko, Yukinari Kato

{"title":"Epitope Mapping of an Anti-Mouse CD39 Monoclonal Antibody Using PA Scanning and RIEDL Scanning.","authors":"Yuki Okada, Hiroyuki Suzuki, Tomohiro Tanaka, Mika K Kaneko, Yukinari Kato","doi":"10.1089/mab.2023.0029","DOIUrl":"10.1089/mab.2023.0029","url":null,"abstract":"A cell-surface ectonucleotidase CD39 mediates the conversion of extracellular adenosine triphosphate into immunosuppressive adenosine with another ectonucleotidase CD73. The elevated adenosine in the tumor microenvironment attenuates antitumor immunity, which promotes tumor cell immunologic escape and progression. Anti-CD39 monoclonal antibodies (mAbs), which suppress the enzymatic activity, can be applied to antitumor therapy. Therefore, an understanding of the relationship between the inhibitory activity and epitope of mAbs is important. We previously established an anti-mouse CD39 (anti-mCD39) mAb, C39Mab-1 using the Cell-Based Immunization and Screening method. In this study, we determined the critical epitope of C39Mab-1 using flow cytometry. We performed the PA tag (12 amino acids [aa])-substituted analysis (named PA scanning) and RIEDL tag (5 aa)-substituted analysis (named RIEDL scanning) to determine the critical epitope of C39Mab-1 using flow cytometry. By the combination of PA scanning and RIEDL scanning, we identified the conformational epitope, spanning three segments of 275-279, 282-291, and 306-323 aa of mCD39. These analyses would contribute to the identification of the conformational epitope of membrane proteins.","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":" ","pages":"44-52"},"PeriodicalIF":0.0,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140177678","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}

引用次数: 0