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用于疾病诊断与治疗的猫 CD19 单克隆抗体的制备方法优化及特异性验证

Optimization of preparation method and specificity verification of cat CD19 monoclonal antibody for disease diagnosis and treatment.

作者信息

Shen Liya, Yan Shuqi, Xu Aoyu, Lan Di, Jiang Xue, Peng Yuehan, Wang Songjun, Wang Zhanzhong, Chen Yongyan

机构信息

Nourse Science Centre for Pet Nutrition, Wuhu, 241200, China.

Shanghai Chowsing Pet Products Co., Ltd, Shanghai, 201702, China.

出版信息

Heliyon. 2024 Jun 15;10(12):e33145. doi: 10.1016/j.heliyon.2024.e33145. eCollection 2024 Jun 30.

DOI:10.1016/j.heliyon.2024.e33145
PMID:39022044
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11253062/
Abstract

CD19 is a surface antigen on B cells that regulates B cell activation and proliferation, participating in B cell signaling. It is expressed in all B cell lineage tumor diseases, making CD19 a significant marker for detecting B cell tumor diseases and an important target for related immunotherapies. In recent years, with the deepening research on canine and feline diseases and the establishment of animal models, the demand for cat CD19 monoclonal antibodies (mAbs) has been steadily increasing. We successfully prepared cat CD19-specific monoclonal antibodies using a KLH-conjugated cat CD19 peptide as an antigen and optimized the antibody production method. The obtained monoclonal antibodies' molecular and cellular affinities were identified using CD19 peptides, eukaryotic overexpressed proteins, and peripheral blood mononuclear cells (PBMCs). The results indicate that the CD19-3H9 and CD19-8A7 monoclonal antibodies prepared in this study specifically bind to the CD19 molecule, demonstrating their suitability for use in ELISA, Western blot, and cell assays. This study successfully produced cat CD19 monoclonal antibodies with specificity and optimized the antibody preparation method, laying the foundation for the diagnosis and targeted drug combination therapy of B cell tumor diseases in both humans and pets.

摘要

CD19是B细胞表面的一种抗原，可调节B细胞的活化和增殖，参与B细胞信号传导。它在所有B细胞谱系肿瘤疾病中均有表达，这使得CD19成为检测B细胞肿瘤疾病的重要标志物以及相关免疫治疗的重要靶点。近年来，随着对犬猫疾病研究的深入以及动物模型的建立，对猫CD19单克隆抗体（mAbs）的需求一直在稳步增加。我们以KLH偶联的猫CD19肽为抗原，成功制备了猫CD19特异性单克隆抗体，并优化了抗体生产方法。使用CD19肽、真核过表达蛋白和外周血单个核细胞（PBMC）鉴定了所获得单克隆抗体的分子亲和力和细胞亲和力。结果表明，本研究制备的CD19 - 3H9和CD19 - 8A7单克隆抗体可特异性结合CD19分子，表明它们适用于ELISA、蛋白质免疫印迹和细胞检测。本研究成功制备了具有特异性的猫CD19单克隆抗体，并优化了抗体制备方法，为人类和宠物B细胞肿瘤疾病的诊断和靶向药物联合治疗奠定了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c6cf/11253062/3ed6fc5caf3e/gr8.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c6cf/11253062/3a59b8b02a34/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c6cf/11253062/3ed6fc5caf3e/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c6cf/11253062/a3685d747b23/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c6cf/11253062/32595746fdd8/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c6cf/11253062/1c050ac4c81d/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c6cf/11253062/2f085069a1b4/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c6cf/11253062/3b6aabde6619/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c6cf/11253062/1f8739e9cb2b/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c6cf/11253062/3a59b8b02a34/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c6cf/11253062/3ed6fc5caf3e/gr8.jpg

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Preparation of a novel monoclonal antibody against active components of PHA-L from Phaseolus vulgaris and its functional characteristics.制备一种针对菜豆 PHA-L 活性成分的新型单克隆抗体及其功能特性。
BMC Biotechnol. 2022 Oct 29;22(1):32. doi: 10.1186/s12896-022-00761-7.
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Comparison of the deep immune profiling of B cell subsets between healthy adults and Sjögren's syndrome.
比较健康成年人和干燥综合征患者 B 细胞亚群的深度免疫特征。
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Anti-CD19 monoclonal antibodies for the treatment of relapsed or refractory B-cell malignancies: a narrative review with focus on diffuse large B-cell lymphoma.抗 CD19 单克隆抗体治疗复发或难治性 B 细胞恶性肿瘤：以弥漫性大 B 细胞淋巴瘤为重点的叙述性综述。
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