title: Two efficient methods for the conjugation of smooth-form lipopolysaccharides with probes bearing hydrazine or amino groups. I. LPS activation with cyanogen bromide.
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.
authors.affiliation: INQUIMAE - DQIAQF, Universidad de Buenos Aires, C1428EHA, Buenos Aires, Argentina. battagli@qi.fcen.uba.ar
authors.forename: Fernando
authors.initials: F
authors.lastname: Battaglini
authors.forename: Diego
authors.initials: D
authors.lastname: Pallarola
mesh_terms: D000588:Amines / Q000737:chemistry*; D002623:Chemistry Techniques, Analytical; D003488:Cyanogen Bromide / Q000737:chemistry*; D003619:Dansyl Compounds / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006735:Horseradish Peroxidase / Q000378:metabolism; D006834:Hydrazines / Q000737:chemistry*; D008050:Lipid A / Q000032:analysis*; D019081:O Antigens / Q000737:chemistry; D019779:Salmonella enterica / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D000588:Amines; D003619:Dansyl Compounds; D006834:Hydrazines; D008050:Lipid A; D019081:O Antigens; C029424:hydrazine; C027913:dansyl hydrazine; D006735:Horseradish Peroxidase; D003488:Cyanogen Bromide