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Abstract:

This chapter presents a conjugation method for coupling probes bearing hydrazine or primary amino groups to a smooth(S)-form lipopolysaccharide (LPS). LPS is modified by the activation of the hydroxyl groups present in its O-antigen moiety with cyanogen bromide in aqueous acetone. The method yields conjugates with good labeling ratios, preserving the endotoxic activity of the lipid A moiety. Conjugation of smooth-form LPS from Salmonella enterica sv. Minnesota with dansyl hydrazine and horseradish -peroxidase yields labeling ratios above 300 nmol dansyl per mg LPS, with nearly no loss of the original endotoxin activity. In the case of horseradish peroxidase, introducing a spacer, a ratio of 28 nmol HRP per mg LPS is obtained, preserving 65% of the original endotoxic activity.

Registro:

Documento: Artículo
Título:Two efficient methods for the conjugation of smooth-form lipopolysaccharides with probes bearing hydrazine or amino groups. I. LPS activation with cyanogen bromide.
Autor:Battaglini, F.; Pallarola, D.
Filiación:INQUIMAE - DQIAQF, Universidad de Buenos Aires, C1428EHA, Buenos Aires, Argentina.
Palabras clave:amine; cyanogen bromide; dansyl chloride; dansyl hydrazine; horseradish peroxidase; hydrazine; hydrazine derivative; lipid A; O antigen; article; chemical analysis; chemistry; mass spectrometry; metabolism; methodology; polyacrylamide gel electrophoresis; Salmonella enterica; Amines; Chemistry Techniques, Analytical; Cyanogen Bromide; Dansyl Compounds; Electrophoresis, Polyacrylamide Gel; Horseradish Peroxidase; Hydrazines; Lipid A; O Antigens; Salmonella enterica; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Año:2011
Volumen:739
Página de inicio:147
Página de fin:160
DOI: http://dx.doi.org/10.1007/978-1-61779-102-4_13
Título revista:Methods in molecular biology (Clifton, N.J.)
Título revista abreviado:Methods Mol. Biol.
ISSN:19406029
CAS:cyanogen bromide, 506-68-3; dansyl chloride, 605-65-2; hydrazine, 10217-52-4, 13775-80-9, 18500-32-8, 302-01-2, 7803-57-8; lipid A, 95991-05-2; Amines; Cyanogen Bromide, 506-68-3; Dansyl Compounds; Horseradish Peroxidase, 1.11.1.-; Hydrazines; Lipid A; O Antigens; dansyl hydrazine, 33008-06-9; hydrazine, 302-01-2
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19406029_v739_n_p147_Battaglini

Citas:

---------- APA ----------
Battaglini, F. & Pallarola, D. (2011) . Two efficient methods for the conjugation of smooth-form lipopolysaccharides with probes bearing hydrazine or amino groups. I. LPS activation with cyanogen bromide. Methods in molecular biology (Clifton, N.J.), 739, 147-160.
http://dx.doi.org/10.1007/978-1-61779-102-4_13
---------- CHICAGO ----------
Battaglini, F., Pallarola, D. "Two efficient methods for the conjugation of smooth-form lipopolysaccharides with probes bearing hydrazine or amino groups. I. LPS activation with cyanogen bromide." Methods in molecular biology (Clifton, N.J.) 739 (2011) : 147-160.
http://dx.doi.org/10.1007/978-1-61779-102-4_13
---------- MLA ----------
Battaglini, F., Pallarola, D. "Two efficient methods for the conjugation of smooth-form lipopolysaccharides with probes bearing hydrazine or amino groups. I. LPS activation with cyanogen bromide." Methods in molecular biology (Clifton, N.J.), vol. 739, 2011, pp. 147-160.
http://dx.doi.org/10.1007/978-1-61779-102-4_13
---------- VANCOUVER ----------
Battaglini, F., Pallarola, D. Two efficient methods for the conjugation of smooth-form lipopolysaccharides with probes bearing hydrazine or amino groups. I. LPS activation with cyanogen bromide. Methods Mol. Biol. 2011;739:147-160.
http://dx.doi.org/10.1007/978-1-61779-102-4_13