Skin reactivity and fibronectin-binding property of TB66 (66-kDa protein of Mycobacterium tuberculosis)
| Autor: | R. G. Navalkar, Rajashri G. Deshpande, D. A. Bhat, Mahfuz Khan |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
Microbiology (medical)
Male medicine.drug_class Guinea Pigs Immunoblotting Molecular Sequence Data Enzyme-Linked Immunosorbent Assay Biology Monoclonal antibody Microbiology Epitope Chromatography Affinity Mycobacterium tuberculosis Affinity chromatography Bacterial Proteins medicine Animals Hypersensitivity Delayed Amino Acid Sequence Bovine serum albumin Sequence Homology Amino Acid Tumor Necrosis Factor-alpha Binding protein Macrophages Antibodies Monoclonal General Medicine biology.organism_classification Fibronectins Fibronectin binding biology.protein Antibody Carrier Proteins |
| Zdroj: | Journal of medical microbiology. 41(6) |
| ISSN: | 0022-2615 |
| Popis: | A 66-kDa protein (TB66) was purified from culture filtrate (CF) and cell sonicate (CS) of Mycobacterium tuberculosis H37Rv by immobilised metal affinity chromatography (IMAC) on a Ni-nitrilotriacetic acid (NTA) column. TB66 was found to be a fibronectin-binding protein as determined by ELISA and could be purified by affinity chromatography with fibronectin-Sepharose. A similar 66-kDa protein could be isolated also from M. bovis, M. bovis BCG, M. africanum and M. tuberculosis H37Ra by IMAC, but not from any other mycobacteria. The NH2-terminal amino-acid sequence of TB66 from H37Rv and M. bovis was identical and showed 85% homology with the N-terminal sequence of bovine serum albumin (BSA). A monoclonal antibody (MAb) OD4AG3 recognised a heat-stable and trypsin-sensitive epitope near the C-terminal end of TB66. This MAb also recognised the 66-kDa protein isolated from the other members of the M. tuberculosis complex. In tests of immunogenicity, TB66 elicited a delayed type hypersensitivity reaction in guinea-pigs immunised with either TB66 or with M. tuberculosis H37Rv. TB66 also elicited an antibody response in immunised guinea-pigs and stimulated murine macrophages to produce tumour necrosis factor. |
| Databáze: | OpenAIRE |
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