title: Purification, characterization and preparation immunomatrixes of S-layer proteins of Thermobifida fusca.
abstract: OBJECTIVE
  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.
  
  
  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.
  
  
  CONCLUSIONS
  It is the first report, and this study shows that the thermophilic S-layer proteins can be used to prepare the immunomatrixes.
  
  
  CONCLUSIONS
  Information in this study can be used to design the strategies for the removal of biologically important materials or toxins from samples.
authors.affiliation: Institute of Biochemistry and Biotechnology, University of the Punjab, Lahore, Pakistan.
authors.forename: S
authors.initials: S
authors.lastname: Pervaiz
authors.forename: T
authors.initials: T
authors.lastname: Shaheen
authors.forename: S
authors.initials: S
authors.lastname: Shaheen
authors.forename: N
authors.initials: N
authors.lastname: Dar
authors.forename: Z Q
authors.initials: ZQ
authors.lastname: Samra
mesh_terms: D000192:Actinomycetales / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D004768:Enterotoxins / Q000276:immunology*; D055767:Immobilized Proteins / Q000737:chemistry; D007158:Immunologic Techniques; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D013211:Staphylococcus aureus / Q000276:immunology
chemical_list: D001426:Bacterial Proteins; D004768:Enterotoxins; D055767:Immobilized Proteins; D008562:Membrane Glycoproteins; C055764:S-layer proteins; C031818:enterotoxin B, staphylococcal
keywords: S-layer protein; SEB toxin; Thermobifida fusca; antibodies; immunomatrixes