Electroeluted outer membrane proteins as immunogens.
Electroeluted outer membrane proteins [(EOmp), (35-37 KDa, porins)] were highly immunogenic in New Zealand White rabbits. An ELISA peak titer of 51,200 to EOmp as compared to 6,400 for non-eluted outer membrane protein (Omp) was demonstrated. EOmp enhanced the antigenicity of Omp possibly due to epitopes which were masked in the non-eluted antigen. Non-eluted, partially purified, Omp elicited high anti-lipopolysaccharide (LPS) titers (25,600); however, electroelution diminished LPS contamination (non detection of LPS chemically and on immunoblots) and greatly reduced the anti-LPS titer (400). It is biologically significant that anti-EOmp antibodies cross-reacted with wild-type urinary pathogens. Specificity for Omp reactivity was demonstrated by ELISA and on immunoblots with absorbed EOmp (LPS-free) antisera. These findings strengthen the rationale for exploring the protective potential of anti-Omp antibodies.
Kawahara
M
M
Department of Urology, Tulane University School of Medicine, New Orleans, Louisiana 70112.
Human
L G
LG
Kawahara
K
K
Domingue
G J
GJ
D000818:Animals; D000942:Antigens, Bacterial / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology* / Q000302:isolation & purification; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000276:immunology; D007074:Immunoglobulin G / Q000096:biosynthesis; D008070:Lipopolysaccharides / Q000276:immunology; D011817:Rabbits
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D007074:Immunoglobulin G; D008070:Lipopolysaccharides