Characterization of monoclonal antibodies for lead-chelate complexes: applications in antibody-based assays.
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.
Zhu
Xiaoxia
X
Key Lab of Monitoring and Management of Plant Diseases and Pests, Ministry of Agriculture, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China.
Hu
Baishi
B
Lou
Yang
Y
Xu
Lina
L
Yang
Fengli
F
Yu
Haini
H
Blake
Diane A
DA
Liu
Fengquan
F
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity; D002614:Chelating Agents / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007118:Immunoassay; D007854:Lead / Q000032:analysis / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D004369:Pentetic Acid / Q000031:analogs & derivatives*
C580995:2-(4-aminobenzyl)diethylenetriaminepentaacetic acid; D000911:Antibodies, Monoclonal; D002614:Chelating Agents; D007854:Lead; D004369:Pentetic Acid