PMID: 11334340May 4, 2001Paper

Method for reducing endotoxin in Moraxella catarrhalis UspA2 protein preparations

Journal of Chromatography. B, Biomedical Sciences and Applications
M J FiskeR Arumugham

Abstract

The UspA2 protein from the bacterium Moraxella catarrhalis is a potential vaccine candidate for preventing human diseases caused by this organism. Before a vaccine can be administered parentally, the level of endotoxin must be reduced as much as possible. However, in this case the endotoxin was very tightly complexed with the UspA2 protein and could not be dissociated with Triton X-100. It was found that it dissociated from the protein with the zwitterionic detergents Zwittergent 3-12 and Zwittergent 3-14. The endotoxin could then be separated from the protein by either ion-exchange or gel filtration chromatography. Using the limulus amoebocyte lysate assay for quantitation, the endotoxin was reduced approximately 20,000-fold. The removal of residual endotoxin from UspA2 preparations had no detrimental effect on the immunological properties of the protein. Mouse antisera raised against UspA2 prior to, and following endotoxin reduction exhibited comparable antibody and bactericidal titers against the tested strains. Further, mice immunized with both preparations, followed by pulmonary challenge with either a homologous or a heterologous isolate, exhibited comparable levels of clearance.

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Citations

Feb 13, 2001·Vaccine·J C McMichael
Aug 1, 2014·Biotechnology Progress·Anne Serdakowski LondonBusola Ruth Alabi
Oct 15, 2009·Biotechnology Progress·Rory E SallachElliot L Chaikof
Jun 24, 2010·Biotechnology Progress·André M LopesAdalberto Pessoa
Feb 18, 2010·Protein Expression and Purification·Yoshihiro FujiwaraKenji Sekikawa
Mar 27, 2015·PloS One·Bożena Szermer-Olearnik, Janusz Boratyński
Jan 25, 2005·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Miao-Fang LinPhilip A Ropp
Oct 20, 2012·Biotechnology Journal·Anne Serdakowski LondonKasey Mackay

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