Production of metal ion-dependent monoclonal antibodies against peptides in bovine prothrombin fragment 1
| Autor: | David G. Klapper, Richard G. Hiskey, Sawsan Hassan Mahassni |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Vitamin
biology Chemistry medicine.drug_class Metal ions in aqueous solution Immunology General Medicine Monoclonal antibody Molecular biology Epitope Metal chemistry.chemical_compound Biochemistry visual_art visual_art.visual_art_medium Phospholipid Binding biology.protein medicine Immunology and Allergy Prothrombin fragment Antibody |
| Zdroj: | Human Antibodies. 17:85-96 |
| ISSN: | 1875-869X 1093-2607 |
| Popis: | Bovine prothrombin fragment 1 (F-1: the amino-terminal 156 residues of prothrombin) is used as a model to study the Ca(II) and phospholipid binding of prothrombin. The 35-46 segment in F-1 posses an α-helical region and three aromatic residues, conserved in several vitamin K-dependent blood coagulation factors. These residues are believed to have a specific function and to be important in the phospholipid binding of F-1. The 47-62 region, a disulfide loop, is believed to stabilize the γ-carboxyglutamic acid domain of the protein. Goals of this research were to produce monoclonal antibodies against the above two sequences, for later functional studies. Antibodies S9-32.8 and S9-5.5 were produced against the 35-46 sequence; antibody S11-23.4 was raised against the 47-62 region. Both S9-32.8 and S9-5.5 bound to F-1 immobilized on ELISA plates in the presence of 10 mM Ca(II) with higher affinity than to F-1 coated in the presence of 10 mM Mg(II) or in the absence of metal ions. S11-23.4 showed greatest binding to F-1 coated in the presence of 10 mM Mg(II). Thus, the epitopes of the antibodies are metal ion-dependent and are developed by Ca(II) binding to F-1. © 2008 - IOS Press and the authors. All rights reserved. |
| Databáze: | OpenAIRE |
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