1. Reactivity of monoclonal antibody E5 with endotoxin. II. Binding to short- and long-chain smooth lipopolysaccharides.
- Author
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Parent JB, Gazzano-Santoro H, Wood DM, Lim E, Pruyne PT, Trown PW, and Conlon PJ
- Subjects
- Animals, Antigens, Bacterial immunology, Binding Sites, Antibody, Binding, Competitive, Escherichia coli chemistry, Escherichia coli immunology, Immunoglobulin M metabolism, Klebsiella chemistry, Lipid A immunology, Mice, O Antigens, Polysaccharides, Bacterial chemistry, Polysaccharides, Bacterial immunology, Proteus chemistry, Pseudomonas chemistry, Radioimmunoassay, Salmonella chemistry, Serratia chemistry, Yersinia chemistry, Antibodies, Monoclonal metabolism, Endotoxins immunology, Gram-Negative Bacteria chemistry, Lipopolysaccharides immunology
- Abstract
The murine monoclonal IgM antibody E5 has been shown to significantly reduce the mortality and morbidity of patients with Gram-negative sepsis in a multicenter randomized placebo-controlled clinical trial. The in vitro binding characteristics of monoclonal antibody (mAb) E5 were studied using highly purified smooth lipopolysaccharide (LPS) isolated from a variety of clinically relevant, wild-type Gram-negative bacteria. Using a sensitive antibody-capture assay which involves immobilized mAb E5 and a chromogenic Limulus amebocyte lysate (LAL) LPS-detection system, mAb E5 was shown to bind to all 15 smooth LPS preparations tested, including LPS isolated from Escherichia, Klebsiella, Proteus, Pseudomonas, Salmonella, Serratia and Yersinia species. When LPS was fractionated according to size by size-exclusion chromatography, mAb E5 was shown to bind to smooth LPS molecules that have long as well as short O-polysaccharide chains. These results confirm and extend those reported previously and demonstrate that the anti-lipid A mAb E5 binds specifically to a diverse spectrum of smooth LPS isolated from wild-type Gram-negative bacteria.
- Published
- 1992