A new rapid and simple method for large-scale purification of mycobacterial lipoarabinomannan.
Lipoarabinomannan (LAM) is a major and structurally important outer cell wall component of all mycobacteria. LAM is also generally regarded as an important immunomodulating substance affecting several immunologic networks and hence important in the pathogenesis of mycobacterial infections. We here describe a new method for large-scale purification of mycobacterial LAM. A crude cell wall preparation was prepared from batch-grown Mycobacterium tuberculosis H37Rv. From this cell wall preparation LAM was purified by sequential extractions and chromatographic steps. From 20 g dry weight cell wall preparation 313 mg of highly purified (> 98%) LAM was obtained in only 3 days. The LAM content of the final purification step was quantified by ELISA using reference LAM as standard. The identity and purity of the LAM preparation was further confirmed by comparison with reference LAM preparation from M. tuberculosis strain Erdman in polyacrylamide gel electrophoresis and Western blots, using reference anti-LAM monoclonals CS-35 and CS-40.
Hamasur
B
B
Swedish Institute for Infectious Disease Control, Stockholm.
Källenius
G
G
Svenson
S B
SB
D000942:Antigens, Bacterial / Q000302:isolation & purification*; D002845:Chromatography / Q000379:methods; D008070:Lipopolysaccharides / Q000302:isolation & purification*; D009169:Mycobacterium tuberculosis / Q000276:immunology*
D000942:Antigens, Bacterial; D008070:Lipopolysaccharides; C050016:lipoarabinomannan