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Abstract:

This chapter presents a conjugation method for coupling probes bearing hydrazine or primary amino groups to a lipopolysaccharide (LPS). LPS is modified by the activation of the hydroxyl groups present in its O-antigen moiety with 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP). 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 110 nmol dansyl/mg LPS, with nearly no loss of the original endotoxic activity. In the case of horseradish peroxidase, introducing a spacer, a ratio of 29 nmol HRP/mg LPS was obtained, preserving 65% of the original endotoxic activity and an enzymatic activity of 120 U/mg.

Registro:

Documento: Artículo
Título:Two efficient methods for the conjugation of smooth-form lipopolysaccharides with probes bearing hydrazine or amino groups. II. LPS activation with a cyanopyridinium agent.
Autor:Battaglini, F.; Pallarola, D.
Filiación:INQUIMAE - DQIAQF, Universidad de Buenos Aires, C1428EHA, Buenos Aires, Argentina.
Palabras clave:1 cyano 4 dimethylaminopyridinium; 1-cyano-4-dimethylaminopyridinium; amine; dansyl chloride; dansyl hydrazine; fluorescent dye; horseradish peroxidase; hydrazine; hydrazine derivative; lipid A; nitrile; O antigen; pyridinium derivative; article; chemistry; mass spectrometry; metabolism; methodology; Salmonella enterica; Amines; Dansyl Compounds; Fluorescent Dyes; Horseradish Peroxidase; Hydrazines; Lipid A; Nitriles; O Antigens; Pyridinium Compounds; Salmonella enterica; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Año:2011
Volumen:739
Página de inicio:161
Página de fin:167
DOI: http://dx.doi.org/10.1007/978-1-61779-102-4_14
Título revista:Methods in molecular biology (Clifton, N.J.)
Título revista abreviado:Methods Mol. Biol.
ISSN:19406029
CAS: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; 1-cyano-4-dimethylaminopyridinium, 59016-55-6; Amines; Dansyl Compounds; Fluorescent Dyes; Horseradish Peroxidase, 1.11.1.-; Hydrazines; Lipid A; Nitriles; O Antigens; Pyridinium Compounds; dansyl hydrazine, 33008-06-9; hydrazine, 302-01-2
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19406029_v739_n_p161_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. II. LPS activation with a cyanopyridinium agent. Methods in molecular biology (Clifton, N.J.), 739, 161-167.
http://dx.doi.org/10.1007/978-1-61779-102-4_14
---------- CHICAGO ----------
Battaglini, F., Pallarola, D. "Two efficient methods for the conjugation of smooth-form lipopolysaccharides with probes bearing hydrazine or amino groups. II. LPS activation with a cyanopyridinium agent." Methods in molecular biology (Clifton, N.J.) 739 (2011) : 161-167.
http://dx.doi.org/10.1007/978-1-61779-102-4_14
---------- MLA ----------
Battaglini, F., Pallarola, D. "Two efficient methods for the conjugation of smooth-form lipopolysaccharides with probes bearing hydrazine or amino groups. II. LPS activation with a cyanopyridinium agent." Methods in molecular biology (Clifton, N.J.), vol. 739, 2011, pp. 161-167.
http://dx.doi.org/10.1007/978-1-61779-102-4_14
---------- VANCOUVER ----------
Battaglini, F., Pallarola, D. Two efficient methods for the conjugation of smooth-form lipopolysaccharides with probes bearing hydrazine or amino groups. II. LPS activation with a cyanopyridinium agent. Methods Mol. Biol. 2011;739:161-167.
http://dx.doi.org/10.1007/978-1-61779-102-4_14