[Preparation and characterization of a monoclonal antibody against human SOCS3].
OBJECTIVE
To prepare and characterize the monoclonal antibody (mAb) against human SOCS3.


METHODS
BALB/c mice were immunized with recombinant GST-SOCS3 protein, from which the spleen cells were isolated and fused with Sp2/0 cells. After several rounds of screening and cloning, the hybridoma cell strain secreting anti-SOCS3 mAb was obtained, whose specificity was evaluated using ELISA and Western blotting, and the titer, immunoglobulin subtype and affinity of the mAb were also measured.


RESULTS
The hybridoma cell strain secreting anti-SOCS3 mAb was identified to belong to IgG2a subtype. The mAb titers in cultural supernatant and acetic fluid were 1:640 and 1:25600, respectively, as determined by ELISA with affinity reaching 4.84x10(6) L/mol.


CONCLUSIONS
The success in anti-SOCS3 mAb preparation provides the basis for further study of the negative regulation of cytokine signal transduction and the immunoregulation in microorganism infections.
Chen
Chang-chun
CC
Department of Microbiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China. chenchangchunchun@163.com
Li
Wan-yi
WY
Zhang
Chong-jie
CJ
Wei
Da-peng
DP
Kuang
Yu
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D011993:Recombinant Fusion Proteins; C106644:SOCS3 protein, human; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins