sepharose and globotriaosylceramide

sepharose has been researched along with globotriaosylceramide* in 2 studies

Other Studies

2 other study(ies) available for sepharose and globotriaosylceramide

Article	Year
Single-step method for purification of Shiga toxin-1 B subunit using receptor-mediated affinity chromatography by globotriaosylceramide-conjugated octyl sepharose CL-4B. Protein expression and purification, 2001, Volume: 22, Issue:2 A new single-step purification method for Shiga toxin (Stx) was developed using receptor-mediated affinity chromatography, in which Gb3Cer (globotriaosylceramide) was conjugated to octyl Sepharose CL-4B as a carrier. This method achieves high yield and high purity in a small column on which Gb3Cer has been immobilized at high density. Using this affinity column, the Stx1 B subunit was purified with homogeneity by a one-step procedure from a crude extract of recombinant Stx1 B subunit-producing Escherichia coli. The purified Stx1 B subunit conserved a natural pentamer structure confirmed by gel filtration and sedimentation equilibrium analysis. Furthermore, the purified Stx1 B subunit was able to bind specifically to Gb3Cer expressed on Burkitt's lymphoma cells. This versatile purification method can be used to isolate various types of natural as well as recombinant Stx, facilitating fundamental studies of human diseases caused by this toxin. Topics: Blotting, Western; Carbohydrate Sequence; Chromatography, Affinity; Chromatography, Gel; Chromatography, Thin Layer; Electrophoresis, Polyacrylamide Gel; Flow Cytometry; Glycolipids; Humans; Molecular Sequence Data; Peptide Fragments; Receptors, Cell Surface; Sepharose; Shiga Toxin 1; Trihexosylceramides; Tumor Cells, Cultured; Ultracentrifugation	2001
An amplification technology for improving sensitivity when measuring components in biological samples. Journal of immunological methods, 1988, Apr-06, Volume: 108, Issue:1-2 A new technology for improving the sensitivity in measuring components in biological samples is described. The method is based on the use of spherical microbeads (detection beads) which contain a large amount of immobilized enzyme and a reagent with biospecific affinity for the component to be detected. These microbeads have been used in a 'sandwich reaction' for visualization of P-fimbriated Escherichia coli which has a known receptor structure (Gal(alpha 1-4)Gal(beta)). In the initial step the bacteria were enriched on a solid support (e.g., a plastic film or beads (greater than 150 microns)) to which the receptor structure had been covalently bound. In the next step detection beads coupled with enzyme and receptor structure were added and finally a chromogenic substrate for the enzyme was used for visualization. A sensitivity of 10(5) bacteria/ml was reached. The detection beads are of general utility and might be useful for the detection of lectins on other pathogens. Topics: Agglutination Tests; Bacterial Adhesion; Enzymes, Immobilized; Escherichia coli; Globosides; Humans; Immunoenzyme Techniques; Latex; Microspheres; Polystyrenes; Pyelonephritis; Sepharose; Serum Albumin, Bovine; Trihexosylceramides	1988

Article

Year

Single-step method for purification of Shiga toxin-1 B subunit using receptor-mediated affinity chromatography by globotriaosylceramide-conjugated octyl sepharose CL-4B.

Protein expression and purification, 2001, Volume: 22, Issue:2

A new single-step purification method for Shiga toxin (Stx) was developed using receptor-mediated affinity chromatography, in which Gb3Cer (globotriaosylceramide) was conjugated to octyl Sepharose CL-4B as a carrier. This method achieves high yield and high purity in a small column on which Gb3Cer has been immobilized at high density. Using this affinity column, the Stx1 B subunit was purified with homogeneity by a one-step procedure from a crude extract of recombinant Stx1 B subunit-producing Escherichia coli. The purified Stx1 B subunit conserved a natural pentamer structure confirmed by gel filtration and sedimentation equilibrium analysis. Furthermore, the purified Stx1 B subunit was able to bind specifically to Gb3Cer expressed on Burkitt's lymphoma cells. This versatile purification method can be used to isolate various types of natural as well as recombinant Stx, facilitating fundamental studies of human diseases caused by this toxin.

Topics: Blotting, Western; Carbohydrate Sequence; Chromatography, Affinity; Chromatography, Gel; Chromatography, Thin Layer; Electrophoresis, Polyacrylamide Gel; Flow Cytometry; Glycolipids; Humans; Molecular Sequence Data; Peptide Fragments; Receptors, Cell Surface; Sepharose; Shiga Toxin 1; Trihexosylceramides; Tumor Cells, Cultured; Ultracentrifugation

2001

An amplification technology for improving sensitivity when measuring components in biological samples.

Journal of immunological methods, 1988, Apr-06, Volume: 108, Issue:1-2

A new technology for improving the sensitivity in measuring components in biological samples is described. The method is based on the use of spherical microbeads (detection beads) which contain a large amount of immobilized enzyme and a reagent with biospecific affinity for the component to be detected. These microbeads have been used in a 'sandwich reaction' for visualization of P-fimbriated Escherichia coli which has a known receptor structure (Gal(alpha 1-4)Gal(beta)). In the initial step the bacteria were enriched on a solid support (e.g., a plastic film or beads (greater than 150 microns)) to which the receptor structure had been covalently bound. In the next step detection beads coupled with enzyme and receptor structure were added and finally a chromogenic substrate for the enzyme was used for visualization. A sensitivity of 10(5) bacteria/ml was reached. The detection beads are of general utility and might be useful for the detection of lectins on other pathogens.

Topics: Agglutination Tests; Bacterial Adhesion; Enzymes, Immobilized; Escherichia coli; Globosides; Humans; Immunoenzyme Techniques; Latex; Microspheres; Polystyrenes; Pyelonephritis; Sepharose; Serum Albumin, Bovine; Trihexosylceramides

1988