Preparement and identification of monoclonal antibody against extracellular region of human CD47 protein
10.3969/j.issn.1673-4130.2015.21.019
- VernacularTitle:CD47分子胞外区蛋白单克隆抗体的制备与鉴定
- Author:
Dongwei LIANG
;
Jihong LIU
- Publication Type:Journal Article
- Keywords:
CD47;
gene expression;
monoclonal antibody
- From:
International Journal of Laboratory Medicine
2015;(21):3125-3127
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare and identify anti‐CD47 monoclonal antibodies .Methods The gene fragment of CD47 was am‐plified by polymerase chain reaction and cloned into prokaryotic expressing vector pET‐32a(+ ) .Purified reconstructed protein was used to immunize BALB/c mice .The immunized spleen cells were isolated and fused with Sp2/0 cells .After screened ,hybridomas secreting anti‐CD47 monoclonal antibody were acquired .Biological activities of antibodies were investigated by Western blot and flow cytometry .Results The recombinant CD47 extracellular domain protein was successfully expressed in BL21 ,and certificated by sodium dodecyl sulfate polyacrylamide gel electropheresis and Western blot .Data of flow cytometry detection demonstrated that the antiserum had high affinity to CD47 protein .Conclusion Recombinant CD47 and its monoclonal antibody ,with high affinity , were successfully prepared ,which could provide reliable tools for the future study of CD47 .
