Use of the Recombinant 38-kDa Antigen of <i>Mycobacterium tuberculosis</i> as an Immunogen for Specific Antisera Production
-
- D'Souza Celine D.
- Radiation Medicine Center, Bhabha Atomic Research Center
-
- Kadival Gururaj V.
- Radiation Medicine Center, Bhabha Atomic Research Center
-
- Samuel Aban M.
- Radiation Medicine Center, Bhabha Atomic Research Center
抄録
The Mycobacterium tuberculosis 38-kDa protein antigen is one of the secreted immunodominant antigens showing high immunogenicity at B-cell and T-cell levels. Although monoclonal antibodies to this antigen have been produced, specific polyclonal antisera is required for standardization of specific immunodiagnostic assays. This protein has been overexpressed and purified from recombinant Escherichia coli using an inducible vector system. During each stage of expression and purification, the recombinant protein was used to immunize mice and rabbits by several methods: 1) as overexpressed protein present as inclusion bodies in recombinant E. coli; 2) embedded in a polyacrylamide gel; 3) fixed to a solid-phase nitrocellulose membrane and 4) emulsified with an adjuvant. All strategies yielded specific antisera as determined by enzyme-linked immunosorbent assay (ELISA) and immunoblot analyses. The results obtained, both quantitative (ELISA) and qualitative (immunoblot) demonstrate that the purified recombinant antigen retains its antigenicity and immunogenicity throughout the various steps in the process of expression and purification and serves as a potent antigen for production of specific antisera to be used in immunoassays.
収録刊行物
-
- MICROBIOLOGY and IMMUNOLOGY
-
MICROBIOLOGY and IMMUNOLOGY 38 (10), 797-800, 1994
Center For Academic Publications Japan
- Tweet
詳細情報 詳細情報について
-
- CRID
- 1570572703128618624
-
- NII論文ID
- 130003484018
-
- ISSN
- 03855600
-
- 本文言語コード
- en
-
- データソース種別
-
- CiNii Articles