Binding Properties of a Monoclonal Antibody Directed toward Lead−Chelate Complexes
Autor: | Haini Yu, M W Brechbiel, J B Delehanty, Mehraban Khosraviani, R C Blake nd, S C Lorbach, Andrey R. Pavlov, Diane A. Blake |
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Rok vydání: | 2000 |
Předmět: |
medicine.drug_class
Stereochemistry Molecular Sequence Data Biomedical Engineering Pharmaceutical Science Bioengineering Ethylenediamine Monoclonal antibody Metal Mice chemistry.chemical_compound Antibody Specificity Organometallic Compounds medicine Animals Chelation Amino Acid Sequence Chelating Agents Immunoassay Pharmacology Mice Inbred BALB C Hybridomas Base Sequence biology Chemistry Pentetic acid Organic Chemistry Antibodies Monoclonal Pentetic Acid Dissociation constant Kinetics Lead visual_art Diethylenetriamine biology.protein visual_art.visual_art_medium Female Antibody Sequence Analysis Biotechnology Nuclear chemistry |
Zdroj: | Bioconjugate Chemistry. 11:267-277 |
ISSN: | 1520-4812 1043-1802 |
DOI: | 10.1021/bc9901548 |
Popis: | A monoclonal antibody (2C12) that recognizes a Pb(II)-cyclohexyldiethylenetriamine pentaacetic acid complex was produced by the injection of BALB/c mice with a Pb(II)-chelate complex covalently coupled to a carrier protein. The ability of purified antibody to interact with a variety of metal-free chelators and metal-chelate complexes was assessed by measuring equilibrium dissociation constants. The antibody bound to metal-free trans-cyclohexyldiethylenetriamine pentaacetic acid (CHXDTPA) with an equilibrium dissociation constant of 2.3 x 10(-)(7) M. Addition of Pb(II) increased the affinity of the antibody for the complex by 25-fold; Pb(II) was the only metal cation (of 15 different di-, tri-, and hexavalent metals tested) that increased the affinity of the antibody for CHXDTPA. The increased affinity was due primarily to an increase in the association rate constant. The antibody also had the ability to interact with ethylenediamine tetraacetic acid (EDTA), diethylenetriamine pentaacetic acid (DTPA), and structurally related derivatives, but with affinities from 50- to 10000-fold less than that determined for CHXDTPA. Addition of metals to EDTA-based chelators reduced the affinity of the antibody for these ligands. However, when DTPA was used as the chelator, addition of Pb(II) increased the affinity of the antibody for the complex by 200-fold. The sensitivity of prototype immunoassays for Pb(II) could be modulated by changing the structure of the immobilized metal-chelate complex and/or the soluble chelator used to complex Pb(II) in the test solution. |
Databáze: | OpenAIRE |
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