Characterization of Monoclonal Antibodies for Lead-Chelate Complexes: Applications in Antibody-Based Assays

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• 作者：Xiaoxia Zhu ; Baishi Hu ; Yang Lou ; Lina Xu ; Fengli Yang ; Haini Yu ; Diane A. Blake ; Fengquan Liu
• 刊名：Journal of Agricultural and Food Chemistry
• 出版年：2007
• 出版时间：June 27, 2007
• 年：2007
• 卷：55
• 期：13
• 页码：4993 - 4998
• 全文大小：83K
• 年卷期：v.55,no.13(June 27, 2007)
• ISSN：1520-5118

文摘

Monoclonal antibodies against lead were generated by immunizing BALB/c mice with lead conjugatedto keyhole limpet hemocyanin (KLH) via a bifunctional chelator, S-2-(4-aminobenzyl)diethylenetriaminepentaacetic acid (DTPA). Stable hybridoma cell lines were produced by fusion of murine splenocytesand SP2/0 myeloma cells. One of the hybridomas generated from this fusion (4/7) synthesized andsecreted an antibody that bound tightly to Pb²⁺-DTPA complexes but not to metal-free DTPA. Theperformance for a competitive inhibition enzyme-linked immunosorbent assay (ELISA) incorporatingthis 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 than0.3% for Cr³⁺, Mn²⁺, Mg²⁺, In³⁺, Ag¹⁺, Ni²⁺, Co²⁺, Zn²⁺, Ca²⁺, Cu¹⁺, and Hg¹⁺. The IC₅₀ value achievedfor lead was 2.72 ± 0.034 M, showing the detection range of 0.092-87.2 M and the lowest detectionlimit of 0.056 ± 0.005 M. Recoveries from the analyte-fortified tap water and ultrapure water werein the range of 80-114% . These results indicate that the ELISA could be a convenient analyticaltool for monitoring lead residues in drinking water.