Production and characterization of rabbit anti-idiotypic antibodies directed against a murine monoclonal anti-B. abortus antibody |
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Authors: | R Adone F Ciuchini R Bianchi G Piccininno C Pistoia |
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Institution: | (1) Istituto Superiore di Sanità, Laboratorio di Medicina Veterinaria, Viale Regina Elena, 299, 00161 Roma, Italia |
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Abstract: | The protective anti-B. abortus monoclonal antibody ISS/32 (Ab1) was used as an immunogen to induce anti-idiotypic antibodies (Ab2) in rabbits. The purpose was to produce and characterize anti-idiotypic antibodies that share conformational similarity with the corresponding bacterial epitope recognized by Ab1. The rabbit anti-IdAb so induced was isolated and affinity-purified. Its specificity for the paratope of Ab1 was determined by evaluating its ability to compete withB. abortus for binding to Ab1 in a competitive ELISA assay. The anti-idiotypic ISS/32 antibodies were able to compete withB. abortus for binding to Ab1 in a dose-dependent manner. Hence, the data indicated that the rabbit anti-Id ISS/32 antibodies reacted with or near the antigen-binding site of Ab1.Abbreviations Ab
antibody
- anti-Id
anti-idiotypic
- ELISA
enzyme linked immunosorbent assay
- IgG
immunoglobulin
- i.p.
intraperitoneal
- mAb
monoclonal antibody
- PBS
phosphate-buffered saline
- s.c.
subcutaneously
- TD
PBS + 0.05% Tween 20% + 1% yeast extract |
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Keywords: | antibody Brucella ELISA epitope idiotype internal-image monoclonal vaccine |
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