Generation of antiserum to specific epitopes.
The ability to prevent disease by immunization with subunit vaccines that incorporate specific epitopes was demonstrated by DiMarchi et al. (1), who used a synthetic peptide to protect cattle against foot-and-mouth disease. However, generation of antibody to peptide antigens is often difficult owing to the small molecular mass and limited chemical complexity. We tested the hypothesis that recombinant DNA and synthetic peptide techniques would make it possible to stimulate vigorous immune responses to specific epitopes of an outer membrane protein of Neisseria gonorrhoeae. The MtrC AP1 sequence from the invariant MtrC gonococcal lipoprotein was genetically fused to maltose binding protein. The resultant fusion protein was used as the primary immunogen to stimulate MtrC AP1-specific antiserum. To enhance antibody production specific to MtrC AP1, boosting immunizations were performed with synthetic MtrC AP1 sequence contained in a multiple antigenic peptide system immunogen. The MtrC AP1-specific antiserum strongly recognized the MtrC protein on Western blots and appeared to bind native MtrC protein in situ. The generation of antibody in this fashion provides the technology to produce antibody to defined epitopes of any protein, including those found in the gonococcal outer membrane. The ability of those antibodies to inhibit bacterial growth or to activate complement protein can then be tested.
Marchion
D C
DC
Division of Biological Sciences, University of Montana, Missoula, USA.
Manning
D S
DS
Shafer
W M
WM
Judd
R C
RC
D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D001426:Bacterial Proteins; D002352:Carrier Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D007106:Immune Sera / Q000276:immunology; D008074:Lipoproteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D058619:Maltose-Binding Proteins; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D026901:Membrane Transport Proteins; D008969:Molecular Sequence Data; D009344:Neisseria gonorrhoeae / Q000276:immunology*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D002352:Carrier Proteins; D000939:Epitopes; D007106:Immune Sera; D008074:Lipoproteins; D058619:Maltose-Binding Proteins; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; C094589:MtrC protein, Neisseria gonorrhoeae; D011993:Recombinant Fusion Proteins