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Two Efficient Methods for the Conjugation of Smooth-Form Lipopolysaccharides with Probes Bearing Hydrazine or Amino Groups. I. LPS Activation with Cyanogen Bromide

 Last Update: 2021-02-16
 Source: Internet
 Author: User

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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.

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