INVESTIGADORES
BATTAGLINI Fernando
capítulos de libros
Título:
Two efficient methods for the conjugation of lipopolysaccharides with probes bearing hydrazine or amino groups. I. LPS activation with cyanogen bromide.
Autor/es:
FERNANDO BATTAGLINI; DIEGO PALLAROLA
Libro:
Bacterial Toxins: Methods and Protocols
Editorial:
Humana Press
Referencias:
Lugar: Totowa, NJ; Año: 2011; p. 161 - 167
Resumen:
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 activation of the hydroxyl groups present in its Oantigen
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 nmoles 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 nmoles HRP per mg LPS is obtained, preserving 65% of the
original endotoxic activity.
Salmonella enterica sv.
Minnesota with dansyl hydrazine and horseradish peroxidase yields labeling
ratios above 300 nmoles 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 nmoles HRP per mg LPS is obtained, preserving 65% of the
original endotoxic activity.