Characterization of monoclonal antibodies for lead-chelate complexes: applications in antibody-based assays.
J Agric Food Chem
; 55(13): 4993-8, 2007 Jun 27.
Article
en En
| MEDLINE
| ID: mdl-17547420
Monoclonal antibodies against lead were generated by immunizing BALB/c mice with lead conjugated to keyhole limpet hemocyanin (KLH) via a bifunctional chelator, S-2-(4-aminobenzyl)diethylenetriamine pentaacetic acid (DTPA). Stable hybridoma cell lines were produced by fusion of murine splenocytes and SP2/0 myeloma cells. One of the hybridomas generated from this fusion (4/7) synthesized and secreted an antibody that bound tightly to Pb2+-DTPA complexes but not to metal-free DTPA. The performance for a competitive inhibition enzyme-linked immunosorbent assay (ELISA) incorporating this antibody was assessed for its sensitivity to changes in pH, ionic strength, and blocking reagents. The cross-reactivities in this ELISA were less than 3% for Fe3+, Cd2+, Hg2+, and Cu2+ and less than 0.3% for Cr3+, Mn2+, Mg2+, In3+, Ag1+, Ni2+, Co2+, Zn2+, Ca2+, Cu1+, and Hg1+. The IC50 value achieved for lead was 2.72 +/- 0.034 microM, showing the detection range of 0.092-87.2 microM and the lowest detection limit of 0.056 +/- 0.005 microM. Recoveries from the analyte-fortified tap water and ultrapure water were in the range of 80-114% . These results indicate that the ELISA could be a convenient analytical tool for monitoring lead residues in drinking water.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Inmunoensayo
/
Quelantes
/
Ácido Pentético
/
Plomo
/
Anticuerpos Monoclonales
Límite:
Animals
Idioma:
En
Revista:
J Agric Food Chem
Año:
2007
Tipo del documento:
Article
País de afiliación:
China
Pais de publicación:
Estados Unidos