Preparation and clinical application of a novel monoclonal antibody against Mycobacterium tuberculosis culture filtrate protein 10
10.3760/cma.j.issn.1674-2397.2013.01.005
- VernacularTitle:抗结核分枝杆菌培养滤液蛋白10的新型单克隆抗体的制备及其临床应用
- Author:
Jianhang LENG
;
Chengmin SHOU
;
Zhigang WU
;
Guangchao ZHUO
;
Keyi WANG
;
Junya SHEN
;
Sheng HUANG
- Publication Type:Journal Article
- Keywords:
Mycobacterium tuberculosis;
Culture filtrate protein 10;
Antigenic peptide;
Antibodies,monoclonal;
Immunologic techniques
- From:Chinese Journal of Clinical Infectious Diseases
2013;(1):23-26
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare a novel monoclonal antibody (mAb) that specifically against Mycobacterium tuberculosis culture filtrate protein 10 (CFP-10).Methods The BALB/c mice were immunized by a peptide with 14 amino acids (aa residues 53 to 66) of CFP-10,and then the splenocytes of mice were fused with myeloma cell line SP2/0.The resultant fused cells were subjected to screening culture,enzyme linked immunosorbent assay (ELISA) assay and subcloning by limited dilution to establish hybridoma cell lines of stable secreting anti-the peptide of CFP-10 antibody.The antibody was purified,and its isotypes were analyzed.Then,the antibody was further evaluated by Western blotting,immunoprecipitation and ELISA in 38 culture supernatant samples of Mycobacterium tuberculosis,20 culture supernatant samples of non-Mycobacterium tuberculosis,32 samples of tuberculous pleural effusion,24 samples of non-tuberculous pleural effusion,and 20 serum samples from healthy controls.Results The isotype of the mAb against the specific peptide of CFP-10 was an IgG1 with κ chain,and it was applicable for Western blotting and immunoprecipitation analysis.ELISA quantitative test showed that the sensitivity and specificity for diagnosis of Mycobacterium tuberculosis were 78.6％ (55/70) and 92.2％ (59/64),respectively.Conclusion The mAb generated against the specific peptide of CFP-10 is high in sensitivity and specificity,and it might be used in the early diagnosis of tuberculosis.
