دورية أكاديمية
Expression and purification of recombinant antigens of Mycobacterium tuberculosis for application in serodiagnosis.
| العنوان: | Expression and purification of recombinant antigens of Mycobacterium tuberculosis for application in serodiagnosis. |
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| المؤلفون: | Kulshrestha A; Department of Biochemistry, University of Delhi South Campus, New Delhi-110 021, India., Gupta A, Verma N, Sharma SK, Tyagi AK, Chaudhary VK |
| المصدر: | Protein expression and purification [Protein Expr Purif] 2005 Nov; Vol. 44 (1), pp. 75-85. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Academic Press Country of Publication: United States NLM ID: 9101496 Publication Model: Print Cited Medium: Print ISSN: 1046-5928 (Print) Linking ISSN: 10465928 NLM ISO Abbreviation: Protein Expr Purif Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Orlando, FL : Academic Press Original Publication: San Diego : Academic Press, c1990- |
| مواضيع طبية MeSH: | Gene Expression*/genetics , Mycobacterium tuberculosis*/genetics , Mycobacterium tuberculosis*/immunology, Antigens, Bacterial/*isolation & purification , Bacterial Proteins/*isolation & purification, Antibodies, Bacterial/immunology ; Antigens, Bacterial/genetics ; Bacterial Proteins/genetics ; Escherichia coli/genetics ; Recombinant Proteins/genetics ; Recombinant Proteins/isolation & purification ; Serologic Tests/methods ; Tuberculosis/diagnosis ; Tuberculosis/immunology ; Tuberculosis/prevention & control |
| مستخلص: | Accurate diagnosis is essential for the treatment, prevention, and control of tuberculosis. Poor specificity of the tuberculin skin test in BCG-vaccinated populations and constraints to implementation of PCR and CMI-based diagnostic assays in developing countries warrant development of easy-to perform robust serological tests. Due to great heterogeneity in humoral response in TB patients, it will be necessary to include several antigens in any diagnostic assay to achieve useful levels of sensitivity and specificity. This needs production of recombinants, soluble versions of mycobacterial antigens in high yields. We have cloned, expressed, and purified a number of mycobacterial proteins in Escherichia coli. This paper describes the expression and purification of four promising sero-reactive proteins namely, ESAT6, CFP10, MTC28, and 14-kDa antigen of Mycobacterium tuberculosis. The protocol involves regulated and slow expression of proteins by using a T7 promoter-based expression vector for obtaining soluble protein followed by a three-step column chromatography procedure employing media with high binding capacity and flow characteristics. The yields of these proteins obtained were several folds higher than previously reported. The purified proteins were useful in detecting antibodies in sera of TB patients (smear positive, smear negative, and extra-pulmonary categories) and in combination with other immunodominant antigens will be useful in increasing the sensitivity to detect M. tuberculosis specific antibodies. |
| المشرفين على المادة: | 0 (Antibodies, Bacterial) 0 (Antigens, Bacterial) 0 (Bacterial Proteins) 0 (Recombinant Proteins) |
| تواريخ الأحداث: | Date Created: 20050629 Date Completed: 20060123 Latest Revision: 20151119 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1016/j.pep.2005.04.014 |
| PMID: | 15982900 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1046-5928 |
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| DOI: | 10.1016/j.pep.2005.04.014 |