Identification of Monoclonal Antibodies against von Willebrand Factor Cleaving Protease (ADAMTS13) and Their Function.

Zhen-ni MA; Jing Ling; Fei Shen; Li-qian Xie; Jie Yin; Jian SU; Chang-geng Ruan

Return

Identification of Monoclonal Antibodies against von Willebrand Factor Cleaving Protease (ADAMTS13) and Their Function.

Author: Zhen-Ni MA ¹ ; Jing LING ² ; Fei SHEN ¹ ; Li-Qian XIE ¹ ; Jie YIN ¹ ; Jian SU ¹ ; Chang-Geng RUAN ³
Author Information

1. Jiangsu Institute of Hematology,Key Laboratory of Thrombosis and Hemostasis of Ministry of Health,The First Affiliated Hospital of Soochow University,Collaborative Innovation Center of Hematology,Soochow University,Suzhou 215000,Jiangsu Province,China.
2. Department of Hematology and Oncology, Children's Hospital of Soochow University, Suzhou 215000, Jiangsu Province,China.
3. Jiangsu Institute of Hematology,Key Laboratory of Thrombosis and Hemostasis of Ministry of Health,The First Affiliated Hospital of Soochow University,Collaborative Innovation Center of Hematology,Soochow University,Suzhou 215000,Jiangsu Province,China. E-mail: ruanchanggeng@suda.edu.cn.
Publication Type:Journal Article
MeSH: ADAMTS13 Protein; Animals; Antibodies, Monoclonal; analysis; immunology; Blotting, Western; Enzyme-Linked Immunosorbent Assay; Hybridomas; Mice; Mice, Inbred BALB C; von Willebrand Factor
From: Journal of Experimental Hematology 2016;24(4):1104-1109
CountryChina
Language:Chinese
Abstract:
OBJECTIVETo construct and identify the monoclonal antibodies against von willebrand factor cleaving protease(ADAMTS13), and to study their biological function.
METHODSBALB/c mice were immunized by purified recombinant ADAMTS13 truncated eukaryotic protein (ADAMTS13-T7). Murine anti-human ADAMTS13 monoclonal antibodies (McAbs) were constructed by standard hybridoma technology and identified by ELISA. The recognition of McAbs with full-length recombinant ADAMTS13 protein was identified by Western blot. In function assay, the influence of McAbs on the proteolysis of vWF by ADAMTS13 was observed.
RESULTSA group of 6 murine anti-ADAMTS13 McAbs was obtained with the clone number 1G11, 2F11, 6G3, 9E1, 10A8 and 10B4. In ELISA, the highest titers of 1G11 and 2F11 were observed, both of which showed a higher affinity to ADAMTS13-T7 than full-length ADAMTS13. The Western blot demonstrated that the 6 McAbs all could recognize ADAMTS13, among which 1G11 and 2F11 showed stronger reaction with ADAMTS13. In addition, under the denatured conditions, 1G11 and 2F11 could inhibit hydrolysis of vWF by ADAMTS13, and that was stronger with the increasing of McAbs concentration.
CONCLUSIONMcAbs against ADAMTS13 have been gained, two of which are inhibitory antibodies.