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.
CITATION STYLE
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. https://doi.org/10.1007/978-1-61779-102-4_14
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