title: Practical applications of high-affinity, albumin-binding proteins from a group G streptococcal isolate.
abstract: Binding proteins that have high affinities for mammalian plasma proteins that are expressed on the surface of bacteria have proven valuable for the purification and detection of several biologically important molecules from human and animal plasma or serum. In this study, we have isolated a high affinity albumin-binding molecule from a group G streptococcal isolate of bovine origin and have demonstrated that the isolated protein can be biotinylated without loss of binding activity and can be used as a tracer for quantification of human serum albumin (HSA). The binding protein can be immobilized and used as a selective capture reagent in a competitive ELISA format using a biotinylated HSA tracer. In this assay format, the sensitivity of detection for 50% inhibition of binding of HSA was less than 1 microg/ml. When attached to the bacterial surface, this binding protein can be used to deplete albumin from human plasma, as analyzed by surface-enhanced laser desorption ionization time of flight mass spectrometry.
authors.affiliation: Department of Biology, Juniata College, 1700 Moore St., Huntingdon, PA 16652, USA.
authors.forename: Emily M
authors.initials: EM
authors.lastname: Coyle
authors.forename: Levi L
authors.initials: LL
authors.lastname: Blazer
authors.forename: Abby A
authors.initials: AA
authors.lastname: White
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Hess
authors.forename: Michael D P
authors.initials: MD
authors.lastname: Boyle
mesh_terms: D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism*; D001710:Biotin / Q000378:metabolism; D006801:Humans; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism*; D013045:Species Specificity; D013291:Streptococcus / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D015415:Biomarkers; D012709:Serum Albumin; D001710:Biotin