Simplified purification of Listeria monocytogenes listeriolysin O and preliminary application in the enzyme-linked immunosorbent assay (ELISA).
A simplified procedure was developed for purification of listeriolysin O (LLO) of Listeria monocytogenes, consisting of hydroxylapatite adsorption chromatography followed by Sepharose S ion exchange chromatography. The LLO (58 kDa) appeared pure in terms of sodium dodecylsulfate polyacrylamide electrophoresis and immunoblots with polyclonal rabbit immune sera. The purified LLO could be stored at -65 degrees C for 1 year without loss of immunoreactivity. Similarly, flat-bottom microtiter strips from two vendors that had been charged with LLO, could be stored at -65 degrees C for up to 3 months without loss of LLO. Three patients with documented listeriosis developed elevated IgG titres against LLO; 2 of the patients revealed minimally raised IgM titres, as determined with the enzyme-linked immunosorbent assay.
Traub
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WH
Institut für Medizinische Mikrobiologie und Hygiene, Universität des Saarlandes, Homburg/Saar, Germany.
Bauer
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D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology; D001426:Bacterial Proteins / Q000276:immunology / Q000302:isolation & purification*; D001427:Bacterial Toxins; D004797:Enzyme-Linked Immunosorbent Assay; D006360:Heat-Shock Proteins / Q000276:immunology / Q000302:isolation & purification*; D006460:Hemolysin Proteins / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D008089:Listeria monocytogenes / Q000276:immunology*; D011817:Rabbits
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; C038705:hlyA protein, Listeria monocytogenes