Recombinant immunotoxins with low endotoxins for clinical and animal studies.
Recombinant immunotoxin (RIT) contains the Fv portion of the antibody fused to the truncated form of toxin and are ongoing in clinical trials for cancer therapy. To obtain high yields of products, RITs are produced in Escherichia coli (E. coli). As the endotoxin came from E. coli cells and is harmful to animals, it is important to produce the RITs with low endotoxin. This section describes the protocols to produce RITs containing low level of endotoxins.
Laboratory of Molecular Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA. OndaM@mail.nih.gov
Masanori
M
Onda
D000818:Animals; D000941:Antigens / Q000276:immunology; D001681:Biological Assay; D016923:Cell Death; D002460:Cell Line; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004731:Endotoxins / Q000096:biosynthesis*; D004926:Escherichia coli / Q000378:metabolism; D005285:Fermentation; D006801:Humans; D000922:Immunotoxins / Q000378:metabolism*; D002479:Inclusion Bodies / Q000378:metabolism; D008070:Lipopolysaccharides / Q000378:metabolism; D008967:Molecular Biology / Q000379:methods*; D010957:Plasmids / Q000378:metabolism; D011489:Protein Denaturation; D058849:Protein Refolding; D011994:Recombinant Proteins / Q000096:biosynthesis*; D012995:Solubility; D014170:Transformation, Genetic
D000941:Antigens; D004731:Endotoxins; D000922:Immunotoxins; D008070:Lipopolysaccharides; D011994:Recombinant Proteins