Engineering and characterization of Hu3A4: A novel humanized antibody with potential as a therapeutic agent against myeloid lineage leukemias

Leukemia stem cells (LSCs) play a critical role in the initiation, recurrence, and resistance to treatment of leukemia. Eradicating LSCs is crucial for the complete elimination of the disease. CD45RA is identified as an important marker for LSC subsets in acute myeloid leukemia (AML), providing a st...

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Main Authors: Sisi Li, Zhujun Wang, Xiaoping Guo, Yongmin Tang
Format: Article
Language:English
Published: Elsevier 2025-01-01
Series:Neoplasia: An International Journal for Oncology Research
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Online Access:http://www.sciencedirect.com/science/article/pii/S1476558624001258
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author Sisi Li
Zhujun Wang
Xiaoping Guo
Yongmin Tang
author_facet Sisi Li
Zhujun Wang
Xiaoping Guo
Yongmin Tang
author_sort Sisi Li
collection DOAJ
description Leukemia stem cells (LSCs) play a critical role in the initiation, recurrence, and resistance to treatment of leukemia. Eradicating LSCs is crucial for the complete elimination of the disease. CD45RA is identified as an important marker for LSC subsets in acute myeloid leukemia (AML), providing a strategic target for therapy. In this report, we introduce Hu3A4, an innovative humanized CD45RA antibody devised to target LSCs expressing this antigen. Hu3A4 retains the antigen-recognition ability of its parental antibody while removing sequences from the variable region that could elicit human anti-mouse immune reactions. The modified variable regions of the heavy and light chains were intricately fused with the constant regions of human IgG1 heavy and light chains, respectively, producing a humanized antibody that emulates the structure of natural IgG. Hu3A4 was produced through recombinant expression in Chinese Hamster Ovary (CHO) cells, which ensured stable gene integration. In vitro tests revealed that Hu3A4 could effectively target and lyse the cells. Further, in vivo studies highlighted Hu3A4′s substantial anti-leukemic activity, significantly prolonging survival times in treated animal models compared to controls (P < 0.01). To summarize, Hu3A4 exhibits remarkable bioactivity and offers a promising therapeutic potential for the treatment of leukemia patients. Progressing Hu3A4 through additional preclinical and clinical studies is crucial to validate its efficacy as a therapeutic agent for leukemia.
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spelling doaj-art-e7843a727c154e5db7836cd6797dfdc62025-08-20T02:49:32ZengElsevierNeoplasia: An International Journal for Oncology Research1476-55862025-01-015910108410.1016/j.neo.2024.101084Engineering and characterization of Hu3A4: A novel humanized antibody with potential as a therapeutic agent against myeloid lineage leukemiasSisi Li0Zhujun Wang1Xiaoping Guo2Yongmin Tang3School of Medicine, Hangzhou City University, #51 Huzhou Street, Hangzhou 310015, ChinaDepartment of Pediatrics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, ChinaDivision of Hematology-Oncology, Pediatric Hematology-Oncology Center, Zhejiang Provincial Research Center for Childhood Leukemia New Diagnostic and Therapeutic Techniques, Children's Hospital, Zhejiang University School of Medicine, National Clinical Medical Research Center for Child Health, #57 Zhugan Road, Yan-an Street, Hangzhou 310006, ChinaDivision of Hematology-Oncology, Pediatric Hematology-Oncology Center, Zhejiang Provincial Research Center for Childhood Leukemia New Diagnostic and Therapeutic Techniques, Children's Hospital, Zhejiang University School of Medicine, National Clinical Medical Research Center for Child Health, #57 Zhugan Road, Yan-an Street, Hangzhou 310006, China; Corresponding author.Leukemia stem cells (LSCs) play a critical role in the initiation, recurrence, and resistance to treatment of leukemia. Eradicating LSCs is crucial for the complete elimination of the disease. CD45RA is identified as an important marker for LSC subsets in acute myeloid leukemia (AML), providing a strategic target for therapy. In this report, we introduce Hu3A4, an innovative humanized CD45RA antibody devised to target LSCs expressing this antigen. Hu3A4 retains the antigen-recognition ability of its parental antibody while removing sequences from the variable region that could elicit human anti-mouse immune reactions. The modified variable regions of the heavy and light chains were intricately fused with the constant regions of human IgG1 heavy and light chains, respectively, producing a humanized antibody that emulates the structure of natural IgG. Hu3A4 was produced through recombinant expression in Chinese Hamster Ovary (CHO) cells, which ensured stable gene integration. In vitro tests revealed that Hu3A4 could effectively target and lyse the cells. Further, in vivo studies highlighted Hu3A4′s substantial anti-leukemic activity, significantly prolonging survival times in treated animal models compared to controls (P < 0.01). To summarize, Hu3A4 exhibits remarkable bioactivity and offers a promising therapeutic potential for the treatment of leukemia patients. Progressing Hu3A4 through additional preclinical and clinical studies is crucial to validate its efficacy as a therapeutic agent for leukemia.http://www.sciencedirect.com/science/article/pii/S1476558624001258CD45RAHumanized antibodyCDR grafting antibodyLeukemic stem cells3A4Targeting therapy
spellingShingle Sisi Li
Zhujun Wang
Xiaoping Guo
Yongmin Tang
Engineering and characterization of Hu3A4: A novel humanized antibody with potential as a therapeutic agent against myeloid lineage leukemias
Neoplasia: An International Journal for Oncology Research
CD45RA
Humanized antibody
CDR grafting antibody
Leukemic stem cells
3A4
Targeting therapy
title Engineering and characterization of Hu3A4: A novel humanized antibody with potential as a therapeutic agent against myeloid lineage leukemias
title_full Engineering and characterization of Hu3A4: A novel humanized antibody with potential as a therapeutic agent against myeloid lineage leukemias
title_fullStr Engineering and characterization of Hu3A4: A novel humanized antibody with potential as a therapeutic agent against myeloid lineage leukemias
title_full_unstemmed Engineering and characterization of Hu3A4: A novel humanized antibody with potential as a therapeutic agent against myeloid lineage leukemias
title_short Engineering and characterization of Hu3A4: A novel humanized antibody with potential as a therapeutic agent against myeloid lineage leukemias
title_sort engineering and characterization of hu3a4 a novel humanized antibody with potential as a therapeutic agent against myeloid lineage leukemias
topic CD45RA
Humanized antibody
CDR grafting antibody
Leukemic stem cells
3A4
Targeting therapy
url http://www.sciencedirect.com/science/article/pii/S1476558624001258
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