Monoclonal antibodies as probes for detecting lipopolysaccharide expression on Escherichia coli from different growth conditions.
Monoclonal antibody (mAb) probes were used to investigate the expression of lipopolysaccharide (LPS) on four Escherichia coli strains, grown under a variety of conditions in batch culture which mimicked some of the in vivo environmental conditions of an infected host. Techniques of silver staining, immunoblotting, whole cell ELISA and flow cytometry were all used to monitor the expression of LPS on the bacteria and the binding of the anti-LPS mAbs. Growth in heat-inactivated sheep serum and magnesium-depleted conditions demonstrated increased expression of LPS core and subsequent increased binding of anti-core mAbs. Magnesium-depleted conditions also resulted in decreased production of O-polysaccharide material. Iron-depleted bacteria showed only minor changes in LPS expression, although increased binding of anti-core mAbs was observed. Nitrogen-deficient/high-carbon conditions, chosen to promote capsule production, resulted in increased expression of O-polysaccharide and decreased binding of anti-core mAbs.
Nelson
D
D
Department of Medical Microbiology, University of Edinburgh Medical School, UK.
Bathgate
A J
AJ
Poxton
I R
IR
D000911:Antibodies, Monoclonal / Q000276:immunology*; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000254:growth & development / Q000276:immunology / Q000378:metabolism*; D005434:Flow Cytometry; D015151:Immunoblotting; D008070:Lipopolysaccharides / Q000032:analysis / Q000096:biosynthesis*; D012834:Silver; D013194:Staining and Labeling
D000911:Antibodies, Monoclonal; D003470:Culture Media; D008070:Lipopolysaccharides; D012834:Silver