Polymyxin B-horseradish peroxidase conjugates as tools in endotoxin research

Anal Biochem. 1992 Dec;207(2):311-6. doi: 10.1016/0003-2697(92)90017-2.

Abstract

The peptide antibiotic Polymyxin B (PMB) binds to bacterial endotoxin (lipopolysaccharide, LPS). We prepared covalent conjugates of PMB and horseradish peroxidase (HRP) by periodation of HRP-linked oligosaccharides followed by direct condensation with PMB. In addition we prepared monoclonal antibodies (Mabs) to PMB. The PMB-HRP conjugates and anti-PMB Mabs were used to study in ELISA the binding of PMB to LPS from Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. In addition, PMB-HRP was used to quantify lipid A in ELISA, and to stain gram-negative bacteria histochemically. For the study of PMB-LPS interaction, PMB-HRP proved to be superior to the anti-PMB Mabs. PMB-HRP conjugates are useful general probes to detect or measure lipid A and LPS of various species using very simple methods and to stain bacteria, and they may obviate the need for many specific antisera. Thus, PMB-HRP conjugates are useful probes for endotoxin research.

MeSH terms

  • Antibodies, Monoclonal*
  • Antigens, Bacterial / analysis*
  • Endotoxins / analysis*
  • Enzyme-Linked Immunosorbent Assay / methods
  • Escherichia coli*
  • Gram-Negative Bacteria / cytology
  • Gram-Negative Bacteria / isolation & purification
  • Horseradish Peroxidase*
  • Immunohistochemistry / methods
  • Klebsiella pneumoniae*
  • Lipid A / analysis*
  • Lipopolysaccharides / analysis*
  • Polymyxin B* / immunology
  • Pseudomonas aeruginosa*

Substances

  • Antibodies, Monoclonal
  • Antigens, Bacterial
  • Endotoxins
  • Lipid A
  • Lipopolysaccharides
  • Horseradish Peroxidase
  • Polymyxin B