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Volume 140, Issue 4

Reactivity of antibodies to heteroclitic peptides based on the major outer-membrane protein Free

Abstract

One problem of peptide vaccines is that antibodies generated against them react poorly with the target sequence on the native protein. Using monoclonal antibodies (mAbs) to the serovar L1 type-specific epitope on the major outer-membrane protein of as our model in conjunction with the Pin Technology Epitope Scanning technique, we had previously identified the critical binding site at this epitope as DAVP. Amino acid substitution showed that AV were essential residues for binding. A series of structurally related (heteroclitic) peptides retaining AV were synthesized. Some of these were found to be much more reactive with the model mAb than peptides of cognate sequence. It was hypothesized that the DAVP peptide only approximated to the conformation of the homologous sequence in the native protein, whereas some of the flexible heteroclitic peptides produced conformations which more closely resembled the native constrained sequence. The key question was whether the most reactive heteroclitic peptide would also generate antibody capable of more efficient binding to the native protein. We therefore immunized mice with one of six heteroclitic peptides or one of two native sequence control peptides. The reactivity of these antisera with the peptide immunogens and with native chlamydial elementary bodies was then evaluated by enzyme immunoassay. Pooled antisera to two of the heteroclitic peptides reacted with significantly greater absorbance ( < 0·05) and at higher dilution with whole chlamydiae than did pooled antisera to the control peptides. This suggests that heteroclitic peptides may in some circumstances be useful to increase the reactivity of site-specific antibodies with epitopes on the native protein important for vaccine development or for serodiagnosis.

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Keyword(s): Chlamydia trachomatis , epitope mapping , major outer membrane protein , peptides and vaccines
© Society for General Microbiology 1994
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1994-04-01
2024-04-24
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Reactivity of antibodies to heteroclitic peptides based on the Chlamydia trachomatis major outer-membrane protein
Microbiology 140, 815 (1994); https://doi.org/10.1099/00221287-140-4-815
/content/journal/micro/10.1099/00221287-140-4-815
/content/journal/micro/10.1099/00221287-140-4-815
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