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Cytotechnology. 2017 Apr;69(2):217-227. doi: 10.1007/s10616-016-0052-5. Epub 2017 Feb 18.

Development and characterization of monoclonal antibody against human IL-37b.

Gao YC1,2, Jia Y1,2, Xiao DQ1, Wang X1,2, Dai YC1,2, Yu SY1,2, Chen C1,2, Zhuang ZG1,2, Fu XX2, Zhang JA1,2, Zheng BY1,2, Chen ZH2, Zhong JX3, Chen ZQ1,2, Xu JF4,5.

Author information

1
Institute of Laboratory Medicine, Guangdong Medical University, No. 1 Xincheng Road, Dongguan, 523808, China.
2
Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics, No. 1 Xincheng Road, Dongguan, 523808, China.
3
Department of Medicine, University of Maryland School of Medicine, Baltimore, MD, 21201, USA.
4
Institute of Laboratory Medicine, Guangdong Medical University, No. 1 Xincheng Road, Dongguan, 523808, China. xujunfa@gdmu.edu.cn.
5
Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics, No. 1 Xincheng Road, Dongguan, 523808, China. xujunfa@gdmu.edu.cn.

Abstract

IL-37 has been described as a natural inhibitor of immune responses. Monoclonal antibody (mAb) against human IL-37b with high affinity and specificity can serve as a molecular probe to detect IL-37 and study IL-37 functions, mechanisms and related signal pathways in inflammatory diseases. However, there are very few such mAbs against human IL-37 commercially available so far. In the current study, monoclonal antibodies against human IL-37b were developed by fusing splenocytes from immunized mouse with SP2/0 myeloma cells and polyethylene glycol. Then the antibodies were screened with prokaryotic expressed human IL-37b protein and eukaryotic expressed human IL-37b protein subsequently. Western blot and flow cytometry analysis revealed that selected mAb clons were able to recognize human IL-37 with high specificity. And more importantly, the IL-37b mAbs were fluorescently labeled and can be directly used in flow cytometry and immunohistochemistry. In conclusion, the current study developed new mAbs against human IL-37b, which are applicable in flow cytometry and immunohistochemistry.

KEYWORDS:

ELISA; Flow cytometry; Hybridoma; Limiting dilution

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