Overpassing an aberrant V(kappa) gene to sequence an anti-idiotypic abzyme with (beta)-lactamase-like activity that could have a linkage with autoimmune diseases

A monoclonal antibody 9G4H9 that exhibits a beta-lactamase-like activity was previously obtained in accordance with the idiotypic network theory. This abzyme presents the most catalytic efficiency in amidase activity described in literature (kcat = 0.9 min-1). Some reports have demonstrated that fun...

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Bibliographic Details
Published in:The FASEB journal 2001-03, Vol.15 (3), p.815-822
Main Authors: Debat, H, Avalle, B, Chose, O, Sarde, C O, Friboulet, A, Thomas, D
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Antibodies, Anti-Idiotypic - genetics
Antibodies, Anti-Idiotypic - metabolism
Antibodies, Catalytic - chemistry
Antibodies, Catalytic - immunology
Antibodies, Catalytic - metabolism
Antibodies, Monoclonal - genetics
Antibodies, Monoclonal - immunology
Antibodies, Monoclonal - metabolism
Base Sequence
Cell Line
Cloning, Molecular
Electrophoresis, Agar Gel
Genes, Immunoglobulin
Immunoglobulin Heavy Chains - genetics
Immunoglobulin kappa-Chains - genetics
Immunoglobulin kappa-Chains - metabolism
Kinetics
Mice
Molecular Sequence Data
Polymerase Chain Reaction - methods
RNA - metabolism
Sequence Alignment
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Summary:A monoclonal antibody 9G4H9 that exhibits a beta-lactamase-like activity was previously obtained in accordance with the idiotypic network theory. This abzyme presents the most catalytic efficiency in amidase activity described in literature (kcat = 0.9 min-1). Some reports have demonstrated that functionality as complex as catalysis may be mimicked in this way. Comparison of the catalytic properties of both enzyme and abzyme previously allowed us to obtain better knowledge about 9G4H9 abzymatic machinery. In attempt to characterize this abzyme, the variable regions of kappa and heavy chain were cloned. We present a 'universal' method to clone the correct Vkappa gene to bypass aberrant Vkappa (abVkappa) produced by MOPC-21-derived hybridomas. Sequences obtained are compared in the GenBank database. The VH and Vkappa genes present some important sequence homology with autoantibodies suggesting a direct relationship between catalytic anti-idiotypic antibody and autoimmunity.
ISSN:0892-6638
DOI:10.1096/fj.00-0410com