An inhibition enzyme-linked immunosorbent assay technique for the detection of endotoxins in proteins extracted from Escherichia coli K12 recombinant DNA.
An inhibition enzyme-linked immunosorbent assay (ELISA) was developed for the specific quantitation of rough (R) mutant E. coli K12 lipopolysaccharide (LPS). Since R-LPS binds poorly to polystyrene microplates, an LPS-BSA covalent complex was prepared following glutaraldehyde activation and used as a coating surface antigen. A 100-fold higher signal was observed using the LPS-BSA complex as solid-phase antigen instead of free LPS. The LPS detection limit obtained was 0.5 ng/ml. This test was applied to hGH extracts produced genetically engineered E. coli K12 and a good correlation was found with the LAL test. This new LPS titration technique will be useful for detecting LPS in complex mixtures and the antigen-antibody reaction will ensure the specificity of the detection.
Peborde
J P
JP
Centre de Transfert en Biotechnologie et Microbiologie (INSA-UPS), Toulouse, France.
Laporte
M
M
Samain
D
D
D000942:Antigens, Bacterial / Q000032:analysis; D001667:Binding, Competitive; D004306:Dose-Response Relationship, Immunologic; D004731:Endotoxins / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000032:analysis*; D013006:Growth Hormone / Q000235:genetics; D008033:Limulus Test; D008070:Lipopolysaccharides / Q000032:analysis*; D011994:Recombinant Proteins / Q000032:analysis*
D000942:Antigens, Bacterial; D004731:Endotoxins; D008070:Lipopolysaccharides; D011994:Recombinant Proteins; D013006:Growth Hormone