Characterization and utility of two monoclonal antibodies to cholera toxin B subunit.
Characterization and utility of two monoclonal antibodies to cholera toxin B subunit.
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DOI：
10.1038/s41598-023-30834-2
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发表时间：
2023-03-15
影响因子：
4.6
通讯作者：
Matoba, Nobuyuki
中科院分区：
文献类型：
--
作者：
Verjan Garcia, Noel;Santisteban Celis, Ian Carlosalberto;Dent, Matthew;Matoba, Nobuyuki
霍乱毒素B亚单位(CTB)是一种有效的免疫调节剂，可用于黏膜疫苗和免疫治疗的开发。为了帮助鉴定CTB及其变异体的多效性生物学功能，我们制备了一组抗CTB的单抗。用酶联免疫吸附试验和表面等离子共振法分析了两株单抗7A12B3和9F9C7与霍乱毒素(CTX)、CTB和具有粘膜愈合活性的重组CTB突变体EPICERTIN的结合亲和力。7A12B3和9F9C7均能与CTX、CTB和EPICERTIN有效结合，在低到亚纳摩尔浓度下具有平衡解离常数，但与大肠杆菌不耐热肠毒素B亚基结合较弱。在Caco 2人结肠上皮细胞的环磷酸腺苷实验中，发现7A12B3单抗对CTX有很强的抑制作用，而9F9C7的抑制活性相对较弱。同时，流式细胞术检测到9F9C7单抗能有效地检测到Caco 2细胞和小鼠脾白细胞表面结合的CTB和EPICERTIN。利用9F9C7免疫组织化学方法，我们证实了口服该蛋白后EPICERTIN在小鼠结肠隐窝中的优先定位。总的来说，这些单抗为研究CTB变异体的生物学功能和临床前发展提供了宝贵的工具。
Cholera toxin B subunit (CTB) is a potent immunomodulator exploitable in mucosal vaccine and immunotherapeutic development. To aid in the characterization of pleiotropic biological functions of CTB and its variants, we generated a panel of anti-CTB monoclonal antibodies (mAbs). By ELISA and surface plasmon resonance, two mAbs, 7A12B3 and 9F9C7, were analyzed for their binding affinities to cholera holotoxin (CTX), CTB, and EPICERTIN: a recombinant CTB variant possessing mucosal healing activity. Both 7A12B3 and 9F9C7 bound efficiently to CTX, CTB, and EPICERTIN with equilibrium dissociation constants at low to sub-nanomolar concentrations but bound weakly, if at all, to Escherichia coli heat-labile enterotoxin B subunit. In a cyclic adenosine monophosphate assay using Caco2 human colon epithelial cells, the 7A12B3 mAb was found to be a potent inhibitor of CTX, whereas 9F9C7 had relatively weak inhibitory activity. Meanwhile, the 9F9C7 mAb effectively detected CTB and EPICERTIN bound to the surface of Caco2 cells and mouse spleen leukocytes by flow cytometry. Using 9F9C7 in immunohistochemistry, we confirmed the preferential localization of EPICERTIN in colon crypts following oral administration of the protein in mice. Collectively, these mAbs provide valuable tools to investigate the biological functions and preclinical development of CTB variants.
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DOI：
10.1016/j.procbio.2020.10.018
发表时间：
2021-03
期刊：
Process biochemistry (Barking, London, England)
影响因子：
--
作者：
Morris DA;Reeves MA;Royal JM;Hamorsky KT;Matoba N
通讯作者：
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影响因子：
168.9
作者：
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通讯作者：
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影响因子：
3.1
作者：
Jobling, MG;Holmes, RK
通讯作者：
Holmes, RK
影响因子：
5.6
作者：
Holmner, Asa;Askarieh, Glareh;Krengel, Ute
通讯作者：
Krengel, Ute
DOI：
10.1007/978-1-61737-957-4_11
发表时间：
2011-01-01
期刊：
PLANT CHROMOSOME ENGINEERING
影响因子：
--
作者：
Matoba, Nobuyuki;Davis, Keith R.;Palmer, Kenneth E.
通讯作者：
Palmer, Kenneth E.