Purification, characterization and preparation immunomatrixes of S-layer proteins of Thermobifida fusca
Article first published online: 14 JUN 2013
© 2013 The Society for Applied Microbiology
Journal of Applied Microbiology
Volume 115, Issue 3, pages 735–743, September 2013
How to Cite
Pervaiz, S., Shaheen, T., Shaheen, S., Dar, N. and Samra, Z.Q. (2013), Purification, characterization and preparation immunomatrixes of S-layer proteins of Thermobifida fusca. Journal of Applied Microbiology, 115: 735–743. doi: 10.1111/jam.12259
- Issue published online: 14 AUG 2013
- Article first published online: 14 JUN 2013
- Accepted manuscript online: 23 MAY 2013 10:43AM EST
- Manuscript Accepted: 18 MAY 2013
- Manuscript Revised: 14 MAY 2013
- Manuscript Received: 20 MAR 2013
- SEB toxin;
- S-layer protein;
- Thermobifida fusca
S-layer proteins are considered as a good nanocarrier due to their binding and self-assembled properties. These can be used to prepare the immunomatrixes for the removal of toxins from the samples.
Methods and Results
Two S-layer proteins 70 and 40 kDa of thermophilic Thermobifida fusca were extracted with guanidine hydrochloride and purified. Antibodies against S-layer proteins were developed, and their monospecificity was checked. Immunogold labelling indicated that these are surface proteins. Immunomatrixes (70-SLIM, 40 SLIM) were prepared by covalently immobilizing S-layer proteins in microwell and further conjugated with anti- Staphylococcus aureus enterotoxin B (SEB) antibodies. The binding of 70 and 40 kDa proteins was observed nearly 7·0 μg cm−1 to binding area, and the conjugation with anti-SEB antibodies was found 1·22 μg μg−1 of 70 kDa and 0·875 μg μg−1 of 40 kDa. The average binding and elution of pure SEB toxin on 70-SLIM and 40-SLIM was 5·0 μg SEB toxin. The SEB toxin in milk samples was also removed on immunomatrixes successfully.
It is the first report, and this study shows that the thermophilic S-layer proteins can be used to prepare the immunomatrixes.
Significance and Impact of Study
Information in this study can be used to design the strategies for the removal of biologically important materials or toxins from samples.