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Abstract:

A conjugation method for coupling probes bearing hydrazine or primary amino groups to a lipopolysaccharide (LPS) is described. LPS is modified through the hydroxyl groups present in its O-antigen moiety by activation with cyanogen bromide in aqueous acetone using triethylamine to enhance the electrophilicity of CNBr. The method yields conjugates with good labeling ratios, preserving the endotoxic activity of the lipid A moiety, which in blood exerts pleiotropic effects on many tissues and organs, resulting in multiple-organ damage, circulatory collapse, and death. Conjugation of smooth-form LPS from Salmonella enterica sv. Minnesota to dansyl hydrazine yielded a labeling ratio of 330 nmol dansyl/mg LPS, with nearly no loss of the original endotoxic activity. In the case of horseradish peroxidase, in which a spacer was introduced, the ratio was 28 nmol HRP/mg of LPS, preserving 65% of the original endotoxic activity. This work shows that under these conditions of CNBr activation, the labeling process has practically no effect on the endotoxic behavior of LPS. The method can be used effectively for the conjugation of LPS to probes bearing primary amino, hydrazine, or hydrazide functional groups. © 2008 Elsevier Inc. All rights reserved.

Registro:

Documento: Artículo
Título:An efficient method for conjugation of a lipopolysaccharide from Salmonella enterica sv. Minnesota with probes bearing hydrazine or amino functional groups
Autor:Pallarola, D.; Battaglini, F.
Filiación:INQUIMAE, Departamento de Química Inorgánica, Analítica y Química Física, Facultad de Ciencias Exactas y Naturales, Ciudad Universitaria-Pabellon 2, C1428EHA Buenos Aires, Argentina
Palabras clave:Cyanogen Bromide; Dansyl hydrazine; Horseradish peroxidase; Lipopolysaccharide conjugates; amino acid; horseradish peroxidase; hydrazine; lipid A; lipopolysaccharide; article; bacterium conjugation; biological activity; death; labeling index; multiple organ failure; nonhuman; priority journal; Salmonella enterica; shock; toxicity; Amines; Chemistry, Analytical; Dansyl Compounds; Electrophoresis; Horseradish Peroxidase; Hydrazines; Molecular Probes; Polyethylene Glycols; Polysaccharides, Bacterial; Salmonella enterica; Spectrometry, Fluorescence; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Armoracia rusticana; Salmonella enterica
Año:2008
Volumen:381
Número:1
Página de inicio:53
Página de fin:58
DOI: http://dx.doi.org/10.1016/j.ab.2008.06.008
Título revista:Analytical Biochemistry
Título revista abreviado:Anal. Biochem.
ISSN:00032697
CODEN:ANBCA
CAS:amino acid, 65072-01-7; hydrazine, 10217-52-4, 13775-80-9, 18500-32-8, 302-01-2, 7803-57-8; lipid A, 95991-05-2; Amines; Dansyl Compounds; dansyl hydrazine, 33008-06-9; Horseradish Peroxidase, EC 1.11.1.-; Hydrazines; Molecular Probes; Polyethylene Glycols; Polysaccharides, Bacterial
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00032697_v381_n1_p53_Pallarola

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Citas:

---------- APA ----------
Pallarola, D. & Battaglini, F. (2008) . An efficient method for conjugation of a lipopolysaccharide from Salmonella enterica sv. Minnesota with probes bearing hydrazine or amino functional groups. Analytical Biochemistry, 381(1), 53-58.
http://dx.doi.org/10.1016/j.ab.2008.06.008
---------- CHICAGO ----------
Pallarola, D., Battaglini, F. "An efficient method for conjugation of a lipopolysaccharide from Salmonella enterica sv. Minnesota with probes bearing hydrazine or amino functional groups" . Analytical Biochemistry 381, no. 1 (2008) : 53-58.
http://dx.doi.org/10.1016/j.ab.2008.06.008
---------- MLA ----------
Pallarola, D., Battaglini, F. "An efficient method for conjugation of a lipopolysaccharide from Salmonella enterica sv. Minnesota with probes bearing hydrazine or amino functional groups" . Analytical Biochemistry, vol. 381, no. 1, 2008, pp. 53-58.
http://dx.doi.org/10.1016/j.ab.2008.06.008
---------- VANCOUVER ----------
Pallarola, D., Battaglini, F. An efficient method for conjugation of a lipopolysaccharide from Salmonella enterica sv. Minnesota with probes bearing hydrazine or amino functional groups. Anal. Biochem. 2008;381(1):53-58.
http://dx.doi.org/10.1016/j.ab.2008.06.008